LOCUS Exported 7713 bp ds-DNA circular SYN 29-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7713)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 29, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7713
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 419..723
/label=P43
misc_feature 756..1088
/label=Apr sp+pro
CDS 1095..1811
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1818..1835
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 2652..2756
/gene="bla"
/label=AmpR promoter
CDS 2757..3617
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3788..4376
/direction=RIGHT
/label=ColE1 ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5779..6549)
/codon_start=1
/label=Kan
/translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
CDS complement(6709..7713)
/codon_start=1
/gene="repB"
/product="RepB replication protein"
/label=repB
/note="from Enterococcus faecalis plasmid pAM-alpha-1"
/db_xref="GI:22652809"
/protein_id="AAN03827.1"
/translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
1 ctatttagtt atttgtttag tcacttataa ctttaacaga tggggttttt ctgtgcaacc
61 aattttaagg gttttcaata ctttaaaaca catacatacc aacacttcaa cgcacctttc
121 agcaactaaa ataaaaatga cgttatttct atatgtatca agataagaaa gaacaagttc
181 aaaaccatca aaaaaagaca ccttttcagg tgcttttttt attttataaa ctcattccct
241 gatctcgact tcgttctttt tttacctctc ggttatgagt tagttcaaat tcgttctttt
301 taggttctaa atcgtgtttt tcttggaatt gtgctgtttt atcctttacc ttgtctacaa
361 accccttaaa aacgttttta aaggctttta agccgtctgt acgttcctta aggaattcga
421 gctcagcttt attgagtgga tgattatatt ccttttgata ggtggtatgt tttcgcttga
481 acttttaaat acagccattg aacatacggt tgatttaata actgacaaac atcaccctct
541 tgctaaagcg gccaaggacg ctgccgccgg ggctgtttgc gtttttgccg tgatttcgtg
601 tattattggt ttacttattt ttttgccaaa gctgtaatgg ctgaaaattc ttacatttat
661 tttacatttt tagaaatggg cgtgaaaaaa agcgcgcgat tatgtaaaat ataaagtgat
721 agcggtacca ttataggtaa gagaggaatg tacacatgaa gaaaccgttg gggaaaattg
781 tcgcaagcac cgcactactc atttctgttg cttttagttc atcgatcgca tcggctgctg
841 aagaagcaaa agaaaaatat ttaattggct ttaatgagca ggaagctgtc agtgagtttg
901 tagaacaagt agaggcaaat gacgaggtcg ccattctctc tgaggaagag gaagtcgaaa
961 ttgaattgct tcatgaattt gaaacgattc ctgttttatc cgttgagtta agcccagaag
1021 atgtggacgc gcttgaactc gatccagcga tttcttatat tgaagaggat gcagaagtaa
1081 cgacaatggg atccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
1141 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
1201 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1261 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1321 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
1381 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
1441 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1501 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
1561 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
1621 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
1681 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
1741 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
1801 agctgtacaa gctcgagcac caccaccacc accacaagct tgtattctat atagtgtcac
1861 ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg ttctaacgac
1921 aatatgtaca agcctaattg tgtagcatct ggcttactga agcagaccct atcatctctc
1981 tcgtaaactg ccgtcagagt cggtttggtt ggacgaacct tctgagtttc tggtaacgcc
2041 gttccgcacc ccggaaatgg tcagcgaacc aatcagcagg gtcatcgcta gccagatcct
2101 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
2161 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
2221 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
2281 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
2341 atgcaggagt cgcataaggg agagcgtcga tatggtgcac tctcagtaca atctgctctg
2401 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
2461 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
2521 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
2581 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
2641 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
2701 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
2761 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
2821 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
2881 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
2941 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
3001 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
3061 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
3121 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
3181 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
3241 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
3301 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
3361 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
3421 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
3481 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
3541 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
3601 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
3661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
3721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
3901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
4021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
4081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
4141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
4201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
4261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
4321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
4381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
4441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
4501 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
4561 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctgatttc
4621 actttttgca ttctacaaac tgcataactc atatgtaaat cgctcctttt taggtggcac
4681 aaatgtgagg cattttcgct ctttccggca accacttcca agtaaagtat aacacactat
4741 actttatatt cataaagtgt gtgctctgcg aggctgtcgg cagtgccgac caaaaccata
4801 aaacctttaa gacctttctt ttttttacga gaaaaaagaa acaaaaaaac ctgccctctg
4861 ccacctcagc aaaggggggt tttgctctcg tgctcgttta aaaatcagca agggacaggt
4921 agtatttttt gagaagatca ctcaaaaaat ctccaccttt aaacccttgc caatttttat
4981 tttgtccgtt ttgtctagct taccgaaagc cagactcagc aagaataaaa tttttattgt
5041 ctttcggttt tctagtgtaa cggacaaaac cactcaaaat aaaaaagata caagagaggt
5101 ctctcgtatc ttttattcag caatcgcgcc cgattgctga acagattaat aatagatttt
5161 agctttttat ttgttgaaaa aagctaatca aattgttgtc gggatcaatt actgcaaagt
5221 ctcgttcatc ccaccactga tcttttaatg atgtattggg gtgcaaaatg cccaaaggct
5281 taatatgttg atataattca tcaattccct ctacttcaat gcggcaacta gcagtaccag
5341 caataaacga ctccgcacct gtacaaaccg gtgaatcatt actacgagag cgccagcttc
5401 atcacttgcc tcccatagat gaatccgaac ctcattacac attagaactg cgaatccatc
5461 ttcatggtga accaaagtga aacctagttt atcgcaataa aaacctatac tctttttaat
5521 atccccgact ggcaatgccg ggatagactg taacattctc acgcataaaa tcccctttca
5581 ttttctaatg taaatctatt accttattat taattcaatt cgctcataat taatcctttt
5641 tcttattacg caaaatggcc cgatttaagc acacccttta ttccgttaat gcgccatgac
5701 agccatgata attactaata ctaggagaag ttaataaata cgtaaccaac atgattaaca
5761 attattagag gtcatcgttc aaaatggtat gcgttttgac acatccacta tatatccgtg
5821 tcgttctgtc cactcctgaa tcccattcca gaaattctct agcgattcca gaagtttctc
5881 agagtcggaa agttgaccag acattacgaa ctggcacaga tggtcataac ctgaaggaag
5941 atctgattgc ttaactgctt cagttaagac cgaagcgctc gtcgtataac agatgcgatg
6001 atgcagacca atcaacatgg cacctgccat tgctacctgt acagtcaagg atggtagaaa
6061 tgttgtcggt ccttgcacac gaatattacg ccatttgcct gcatattcaa acagctcttc
6121 tacgataagg gcacaaatcg catcgtggaa cgtttgggct tctaccgatt tagcagtttg
6181 atacactttc tctaagtatc cacctgaatc ataaatcggc aaaatagaga aaaattgacc
6241 atgtgtaagc ggccaatctg attccacctg agatgcataa tctagtagaa tctcttcgct
6301 atcaaaattc acttccacct tccactcacc ggttgtccat tcatggctga actctgcttc
6361 ctctgttgac atgacacaca tcatctcaat atccgaatag ggcccatcag tctgacgacc
6421 aagagagcca taaacaccaa tagccttaac atcatcccca tatttatcca atattcgttc
6481 cttaatttca tgaacaatct tcattctttc ttctctagtc attattattg gtccattcac
6541 tattctcatt cccttttcag ataattttag atttgctttt ctaaataaga atatttggag
6601 agcaccgttc ttattcagct attaataact cgtcttccta agcatccttc aatcctttta
6661 ataacaatta tagcatctaa tcttcaacaa actggcccgt ttgttgaact actctttaat
6721 aaaataattt ttccgttccc aattccacat tgcaataata gaaaatccat cttcatcggc
6781 tttttcgtca tcatctgtat gaatcaaatc gccttcttct gtgtcatcaa ggtttaattt
6841 tttatgtatt tcttttaaca aaccaccata ggagattaac cttttacggt gtaaaccttc
6901 ctccaaatca gacaaacgtt tcaaattctt ttcttcatca tcggtcataa aatccgtatc
6961 ctttacagga tattttgcag tttcgtcaat tgccgattgt atatccgatt tatatttatt
7021 tttcggtcga atcatttgaa cttttacatt tggatcatag tctaatttca ttgccttttt
7081 ccaaaattga atccattgtt tttgattcac gtagttttct gtattcttaa aataagttgg
7141 ttccacacat accaatacat gcatgtgctg attataagaa ttatctttat tatttattgt
7201 cacttccgtt gcacgcataa aaccaacaag atttttatta atttttttat attgcatcat
7261 tcggcgaaat ccttgagcca tatctgacaa actcttattt aattcttcgc catcataaac
7321 atttttaact gttaatgtga gaaacaacca acgaactgtt ggcttttgtt taataacttc
7381 agcaacaacc ttttgtgact gaatgccatg tttcattgct ctcctccagt tgcacattgg
7441 acaaagcctg gatttacaaa accacactcg atacaacttt ctttcgcctg tttcacgatt
7501 ttgtttatac tctaatattt cagcacaatc ttttactctt tcagcctttt taaattcaag
7561 aatatgcaga agttcaaagt aatcaacatt agcgattttc ttttctctcc atggtctcac
7621 ttttccactt tttgtcttgt ccactaaaac ccttgatttt tcatctgaat aaatgctact
7681 attaggacac ataatattaa aagaaacccc cat
//