LOCUS Exported 5216 bp ds-DNA circular SYN 29-JUN-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBiFC-VN173
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5216)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sunday, September 4, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5216
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 141..157
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
enhancer 318..697
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 698..901
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 935..958
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
CDS 1028..1546
/codon_start=1
/note="VN173"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
polyA_signal 1557..2179
/note="hGH poly(A) signal"
/note="human growth hormone polyadenylation signal"
promoter 2208..2537
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2388..2523
/note="SV40 ori"
/note="SV40 origin of replication"
promoter complement(2576..2594)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2608..2624)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2632..2648
/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(2656..2686)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2701..2722
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3010..3598)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3769..4629)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4630..4734)
/gene="bla"
/note="AmpR promoter"
rep_origin 4761..5216
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
961 tgcggccgcg aattcatcga tagatctgat atcggtacca gtcgactcta gaagatccat
1021 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
1081 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
1141 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
1201 gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca
1261 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
1321 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
1381 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
1441 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
1501 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgagtagg gatcccgggt
1561 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
1621 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
1681 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
1741 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
1801 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
1861 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
1921 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
1981 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
2041 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
2101 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
2161 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
2221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
2341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
2401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
2461 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
2521 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
2581 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
2641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
2701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
2761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
2821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
3421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
3721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
3781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
3841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
3901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
3961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
4021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
4081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
4141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
4201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
4261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
4321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
4381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
4441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
4501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
4561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
4621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
4681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
4741 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
4801 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
4861 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
4921 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
4981 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
5041 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
5101 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
5161 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//