LOCUS       V041759                 8548 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041759
VERSION     V041759
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..4038
                     /label="Itgb5(NM_001145884)"
                     /note="Itgb5(NM_001145884)"
                     /gene="Itgb5"
     polyA_signal    4115..4339
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4385..4813
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4827..5156
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5223..6017
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6191..6312
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6361..6377)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6385..6401
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6409..6439)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6454..6475
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6763..7348)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7519..8379)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8380..8484)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgccgcgg gtgcccgcga ccctctacgc ctgtctgctc
     1681 gggctctgcg cgctcgttcc gcgcctcgca gggctcaaca tatgcactag tggaagtgcc
     1741 acctcgtgtg aagaatgcct gttgatccac ccaaaatgtg cctggtgctc caaagagtac
     1801 tttggcaatc cacggtccat cacctctcgg tgtgacctga aggcaaacct catccggaat
     1861 ggctgtgaag gtgagattga gagtccagcc agcagcaccc acgtcctccg gaacctacct
     1921 ctcagcagca agggttccag tgccacgggc tctgacgtca tccagatgac gccgcaggag
     1981 attgcagtga gcctccggcc aggcgagcag actacgttcc agctgcaggt gcgccaggtg
     2041 gaggactacc ctgtagacct gtactacctg atggacctct ccctctccat gaaggatgac
     2101 ttggagaaca tccggagcct gggcaccaag cttgcggagg aaatgaggaa gctcactagt
     2161 aacttccgct taggtttcgg gtcttttgtt gacaaggaca tctctccttt ctcctacacg
     2221 gcaccgagat accagaccaa tccgtgtatt ggttacaagt tattccccaa ctgcgtcccc
     2281 tccttcgggt tccggcatct gctgcctctc acagacagag tcgacagctt caacgaggaa
     2341 gtgaggaagc agagggtgtc ccggaaccga gatgcccccg agggggggtt tgatgcggtc
     2401 ctccaggctg ctgtctgcaa ggagaagatc ggatggcgaa aagatgctct gcacttgctg
     2461 gtgttcacaa cagacgatgt gccccacatc gcactggatg gaaaactggg tggcctggtc
     2521 cagccccacg atggccagtg tcacctgaat gaagccaatg agtacacagc ctctaaccag
     2581 atggactatc catcgcttgc cttgcttggg gagaagctgg cagagaacaa tatcaacctc
     2641 atttttgctg tgacgaagaa ccactatatg ctctacaaga attttacagc cctgatacct
     2701 ggaaccactg tggagatttt gcatggagat tccaaaaata ttattcaact gattatcaat
     2761 gcgtacagta gcatccgggc taaagtggag ctgtcagtgt gggatcagcc agaagacctt
     2821 aatctcttct tcactgccac ctgccaagat ggcatatctt accctggtca gaggaagtgt
     2881 gagggtctga agattgggga cacggcatcc tttgaagtgt ccgtggaggc tcggagctgc
     2941 cccggcagac aagcagcaca gtctttcacc ttgaggcccg tgggcttccg ggacagtctg
     3001 caggtggaag tcgcctacaa ttgcacatgc ggctgtagca cggggctgga gcccaacagt
     3061 gccagatgca gtgggaatgg aacatacacc tgtgggctgt gcgagtgtga ccccggctac
     3121 ctgggcacta ggtgcgagtg ccaggagggg gagaaccaga gcgggtacca gaacctgtgc
     3181 cgggaggcag agggcaagcc tctgtgcagc gggcgtggag agtgtagctg caaccagtgc
     3241 tcctgcttcg agagtgagtt cgggaggatc tacggacctt tctgcgagtg tgacagcttt
     3301 tcctgtgcca gaaacaaggg cgtcctatgc tcaggccatg gagagtgtca ctgtggagaa
     3361 tgcaaatgcc acgcaggtta cattggggac aattgtaact gctcaacaga cgtcagcaca
     3421 tgcagggcca aggatgggca gatctgcagt gaccgaggcc gttgtgtctg tgggcagtgc
     3481 cagtgcacag agcctggagc ctttggggag acgtgtgaga agtgcccaac ctgcccggat
     3541 gcttgcagct ctaagagaga ctgtgtcgaa tgcttgctac ttcaccaggg gaaacctgac
     3601 aaccagacct gccaccacca gtgcaaagat gaggtgatca cgtgggtaga caccatcgtc
     3661 aaagatgacc aggaggctgt gctttgcttc tacaaaactg ctaaggactg cgttatgatg
     3721 ttcagctaca cagaactgcc caatgggagg tccaacttga cggtcctccg ggagccagaa
     3781 tgtggaagtg cccccaatgc catgaccatc ctgctggctg tggttggcag catcctcctg
     3841 attgggatgg cactcctggc catctggaag ctgctcgtca ccatccacga ccgccgagag
     3901 tttgccaagt tccaaagtga gcgctccagg gcccgttatg aaatggcctc aaaccccctg
     3961 tacagaaagc ccatctccac acacactgtc gatttcgcct tcaacaagtt caacaaatcc
     4021 tacaatggct cagtggactg aggatcctga tatccagcac agtggcggcc gctcgagtct
     4081 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4141 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4201 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4261 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4321 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4381 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5041 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5101 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5161 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5401 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5521 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6001 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6061 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6121 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6181 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6301 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     7321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8521 atcgggagat ctcccgatcc cctatggt
//
