LOCUS Exported 7509 bp ds-DNA circular SYN 12-JAN-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pOTENT-CMV-Flag-His-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7509)
AUTHORS Huanghua
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7509
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 21..123
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 211..361
/note="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
enhancer 419..722
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 723..926
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 954..972
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1073..1096
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
CDS 1106..1123
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1294..1415
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2342..2671
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2522..2657
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2729..3328
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 3329..3553
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
repeat_region 4464..4566
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
rep_origin complement(4838..5426)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 6258..7052
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
rep_origin complement(7100..7480)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 ttcgaattta aattaattaa catcatcaat aatatacctt attttggatt gaagccaata
61 tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg
121 tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg
181 tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg
241 gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc
301 gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagcgcgtaa
361 tactgtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
601 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
661 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
721 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
841 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
901 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagtaatacg
961 actcactata gggagaggat ccggtaccga ggagatctgc cgccgcgatc gccggcgcgc
1021 cagatctcaa gcttaactag ctagcggacc gacgcgtacg cggccgctcg aggattataa
1081 ggatgacgac gataaattcg tcgagcacca ccaccaccac cactaataag gtttatccga
1141 tccaccggat ctagataaga tatccgatcc accggatcta gataactgat cataatcagc
1201 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
1261 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
1321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
1381 agttgtggtt tgtccaaact catcaatgta tcttaacgcg gatctgggcg tggttaaggg
1441 tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg
1501 ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc
1561 gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc
1621 ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg
1681 agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg
1741 actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg
1801 acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt
1861 ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca
1921 atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt
1981 cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt
2041 cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat
2101 acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg
2161 gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt
2221 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt
2281 taagctggga tgggtgcata cgtgtttaaa ctagggataa cagggtaatc ggctgtggaa
2341 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
2401 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
2461 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
2521 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
2581 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
2641 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
2701 cggatctgat caagagacac gtacgaccat gaccgagtac aagcccacgg tgcgcctcgc
2761 cacccgcgac gacgtccccc gggcagtacg caccctcgcc gccgcgttcg ccgactaccc
2821 cgccacgcgc cacaccgtcg atccagaccg ccacatcgag cgggtcaccg agctgcaaga
2881 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
2941 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
3001 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
3061 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
3121 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
3181 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
3241 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
3301 gtgcatgacc cgcaagcccg gtgcctgact gtgccttcta gttgccagcc atctgttgtt
3361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
3421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
3481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
3541 gtgggctcta tggtagggat aacagggtaa tgtttaaact ataaaatgca aggtgctgct
3601 caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca tcgtagtcat gctcatgcag
3661 ataaaggcag gtaagctccg gaaccaccac agaaaaagac accatttttc tctcaaacat
3721 gtctgcgggt ttctgcataa acacaaaata aaataacaaa aaaacattta aacattagaa
3781 gcctgtctta caacaggaaa aacaaccctt ataagcataa gacggactac ggccatgccg
3841 gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca ccaccgacag ctcctcggtc
3901 atgtccggag tcataatgta agactcggta aacacatcag gttgattcac atcggtcagt
3961 gctaaaaagc gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt
4021 acagccccca taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa
4081 aaaccctcct gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttcc
4141 acagcggcag ccataacagt cagccttacc agtaaaaaag aaaacctatt aaaaaaacac
4201 cactcgacac ggcaccagct caatcagtca cagtgtaaaa aagggccaag tgcagagcga
4261 gtatatatag gactaaaaaa tgacgtaacg gttaaagtcc acaaaaaaca cccagaaaac
4321 cgcacgcgaa cctacgccca gaaacgaaag ccaaaaaacc cacaacttcc tcaaatcgtc
4381 acttccgttt tcccacgtta cgtcacttcc cattttaaga aaactacaat tcccaacaca
4441 tacaagttac tccgccctaa aacctacgtc acccgccccg ttcccacgcc ccgcgccacg
4501 tcacaaactc caccccctca ttatcatatt ggcttcaatc caaaataagg tatattattg
4561 atgatgttaa ttaatttaaa ttcgaacatg catggatcca tatgcggtgt gaaataccgc
4621 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
4681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
4741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
4801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
4861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
4921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
4981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
5041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
5101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
5161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
5221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
5281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
5341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
5401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
5461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
5521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
5581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
5641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
5701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
5761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
5821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
5881 ttaatagttt gcgcaacgtt gttgccattg ctgcagccat gagattatca aaaaggatct
5941 tcacctagat ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
6001 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
6061 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
6121 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
6181 tggctttctt gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag
6241 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
6301 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
6361 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
6421 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
6481 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
6541 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
6601 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
6661 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
6721 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
6781 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
6841 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
6901 ccgaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
6961 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
7021 ccttctatcg ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc
7081 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
7141 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
7201 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
7261 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
7321 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
7381 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
7441 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggatcg
7501 aattaattc
//