LOCUS       V029952                 7750 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029952
VERSION     V029952
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..3240
                     /label="Tbxas1(NM_011539)"
                     /note="Tbxas1(NM_011539)"
                     /gene="Tbxas1"
     polyA_signal    3317..3541
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3587..4015
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4029..4358
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4425..5219
                     /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    5393..5514
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5563..5579)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5587..5603
                     /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(5611..5641)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5656..5677
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5965..6550)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6721..7581)
                     /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(7582..7686)
                     /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 catggaagtg ttggggcttc tcaagtttga agtcagtggt
     1681 accatagtga ctgtgactct gctcgtggct ctcttggccc tcctgaaatg gtactccatg
     1741 tcagctttct caagactgga gaagttgggc atcaggcacc ccaagccttc tccttttgtt
     1801 ggaaacttga tgtttttccg ccagggtttt tgggagagcc aattggaact ccgagagcga
     1861 tacgggcctc tgtgtgggta ctatcttggc cgtcggatgc acgttgtcat ttcagagcca
     1921 gacatgatca agcaggtgtt ggttgagaac ttcagtaact tttccaacag aatggcctca
     1981 ggtctggaac ccaagatggt agcagacagt gtcctgttgt tacgtgacag aagatgggag
     2041 gaagtcaggg gtgccctgat gtcttcattc agtcctgaaa agttggatga gatgacacct
     2101 ctcatcagcc aagcctgtga acttctcgtg gctcacttaa aacgctatgc agcatccagg
     2161 gacgcattca acatccagag gtgttactgc tgttacacca tagatgtggt ggccagtgtg
     2221 gcctttggca cccaggtgga ctcccagaat tctccagaag atccctttgt gcaacactgc
     2281 cggcgtgctt ccaccttctg tatccccagg cctctcctgg tattaatctt atcatttcca
     2341 tccataatgg tcccattggc ccggattctg cccaataaga accgagatga actgaatggc
     2401 ttttttaaca cactcattag gaatgtgatt gccttacggg accagcaagc agcagaagag
     2461 aggcggagag acttcctgca gatggtgctg gatgcccagc actccatgaa ctctgtgggc
     2521 gtggaaggct ttgacatggt cccagaatcc ctgtcctctt ctgagtgcac aaaggaacca
     2581 ccccaaaggt gccatcctac ctccacatct aagcctttca ctgtggatga aattgtgggc
     2641 caggccttcc tcttcctcat tgcgggccat gaggtcatca caaacacgct gtccttcatc
     2701 acatacctgc tggccaccca ccctgactgc caggagaggc ttctgaaaga ggtggacctc
     2761 ttcatgggga agcacccagc ccctgagtac cacagcctgc aggaaggtct gccgtatctg
     2821 gacatggtga tttcagagac cctgaggatg tacccaccag ctttcaggtt cacacgggag
     2881 gcagcacagg actgtgaggt gctgggacaa cgtatccctg caggtacagt gctggagata
     2941 gctgtgggtg ccctacacca tgacccagag cactggccga atcctgagac ctttgaccct
     3001 gaaaggttca cagcagaggc ccggcttcag cggaggccgt tcacatacct gccctttgga
     3061 gctggcccca ggagctgcct cggagtgcgg ctgggcctgc tggtggtcaa gctgacaata
     3121 ctccaggtcc tacacaagtt ccgctttgaa gccagccctg agactcaggt tccacttcag
     3181 ctagaatcca aatctgccct aggccccaaa aatggagtct acatcaagat tgtgtcacgc
     3241 tgaggatcct gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3301 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3361 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3421 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3481 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3541 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3601 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3661 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3721 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3781 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3841 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3901 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3961 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4261 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4381 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4441 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4501 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4561 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4621 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4681 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4741 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4801 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4861 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4921 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4981 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5041 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5101 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5161 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5221 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5281 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5341 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5401 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5461 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5521 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5581 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5641 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5701 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5761 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5821 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5881 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5941 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6001 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6061 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6481 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7681 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7741 cccctatggt
//
