LOCUS Exported 8895 bp ds-DNA circular SYN 14-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8895)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8895
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 82..106
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS complement(166..399)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
primer_bind 309..325
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 329..385
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(395..411)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 419..435
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(443..473)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 488..509
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 700..1377
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 1440..2237
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 2294..2468
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 2546..2570
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
CDS 3098..3757
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
rep_origin 4028..4616
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 4802..4942
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin 5286..5480
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(5546..6619)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(7048..7677)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
1 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgttattcta
61 ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt
121 aaactgaagg cgggaaacga caatctgatc caagctcaag ctgctctagc attcgccatt
181 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
241 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
301 acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg actctagagg
361 atccccgggt accgagctcg aattcgtaat catggtcata gctgtttcct gtgtgaaatt
421 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
481 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
541 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
601 tgcgtattgg ctagagcagc ttgccaacat ggtggagcac gacactctcg tctactccaa
661 gaatatcaaa gatacagtct cagaagacca aagggctatt gagacttttc aacaaagggt
721 aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca tcaaaaggac
781 agtagaaaag gaaggtggca cctacaaatg ccatcattgc gataaaggaa aggctatcgt
841 tcaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt
901 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg ataacatggt
961 ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag
1021 ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc
1081 agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca
1141 tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga
1201 tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa
1261 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
1321 ttcgcaagac cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgaaat
1381 caccagtctc tctctacaaa tctatctctc tcgagctttc gcagatctgt cgatcgacca
1441 tggggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
1501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
1561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
1621 tccaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
1681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
1741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
1801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
1861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
1921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
1981 acggcgagga tctcgtcgtg acacatggcg atgcctgctt gccgaatatc atggtggaaa
2041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
2101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
2161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
2221 ttgacgagtt cttctgagcg ggactctggg gttcggatcg atcctctagc tagagtcgat
2281 cgacaagctc gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg
2341 ttcctatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
2401 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
2461 tccagatccc ccgaattaat tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt
2521 tattaagttg tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc
2581 aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag
2641 tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga
2701 tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag
2761 ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcaat tcgggcacga
2821 acccagtgga cataagcctc gttcggttcg taagctgtaa tgcaagtagc gtaactgccg
2881 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
2941 ttcatggctt cttgttatga catgtttttt tggggtacag tctatgcctc gggcatccaa
3001 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
3061 gcagcagggc agtcgcccta aaacaaagtt aaacatcatg gagaaaaaaa tcactggata
3121 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
3181 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
3241 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
3301 tgctcatccg gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
3361 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
3421 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
3481 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
3541 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
3601 ccccgttttc acaatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
3661 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
3721 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
3781 cgatcctcta gctagaaatt cgttcaagcc gacgccgctt cgccggcgtt aactcaagcg
3841 attagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag tctgctttta
3901 ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca tgcatgacca
3961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
4141 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
4201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
4261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
4321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
4381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
4441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
4501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
4561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
4621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
4681 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
4741 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
4801 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
4861 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
4921 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
4981 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
5041 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc
5101 gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg cctggccgta
5161 ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg gcggggcgta
5221 gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg cgctggccag
5281 acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa gagttttagg
5341 cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac cggttcccaa
5401 tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc tttgggttcc
5461 caatgtacgt gctatccaca ggaaagagac cttttcgacc tttttcccct gctagggcaa
5521 tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc tcgatcaggt
5581 tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc aaatcgtact
5641 ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc ttgaactctc
5701 cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct gccttgcctg
5761 cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat
5821 cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg tacatccaat
5881 cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg atcttgtagc
5941 ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg gccttcttcg
6001 tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc aggtcgtctt
6061 tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg tgtggacgga
6121 acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat tcggttagat
6181 gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg gccggccctg
6241 cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca gctcgtcggt
6301 cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg cgggtgccca
6361 cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa
6421 tcgacggcgc accggctgcc ggcggttgcc gggattcttt gcggattcga tcagcggccg
6481 cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg
6541 cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt accgggccgg
6601 atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc cattgcaggg
6661 ccggcagaca acccagccgc ttacgcctgg ccaaccgccc gttcctccac acatggggca
6721 ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt tagccgctaa
6781 aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg atgtattcag
6841 atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc ttggtgtgat
6901 cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc aggctggcca
6961 acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt
7021 tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt cagcggccag
7081 cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc
7141 ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc
7201 gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga tcgcccgcga
7261 cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct taaccagctc
7321 caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa tcagcacgaa
7381 gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc cgtcgatcac
7441 tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc
7501 gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg
7561 atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg ctagatgggt
7621 tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa ccttcatgcg
7681 ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac cgcatgacgc
7741 aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt ttcttcagcg
7801 gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag gatttcatgc
7861 agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc gatcatctcc
7921 gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg
7981 cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt
8041 cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa
8101 gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct ttcacggtgc
8161 ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc
8221 gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga
8281 tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg
8341 ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg ccggcctcct
8401 ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg
8461 tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc tccgggcggt
8521 cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt
8581 cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag cccagcaggc
8641 cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa gtattctact
8701 ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc
8761 gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact gaacgtcaga
8821 agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc cgaggggaac
8881 cctgtggttg gcatg
//