LOCUS       V022863                 6870 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022863
VERSION     V022863
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             884..1600
                     /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             1619..2359
                     /label="STC1(NM_003155)"
                     /note="STC1(NM_003155)"
                     /gene="STC1"
     polyA_signal    2437..2661
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2707..3135
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3149..3478
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3545..4339
                     /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    4513..4634
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4683..4699)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4707..4723
                     /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(4731..4761)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4776..4797
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5085..5670)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5841..6701)
                     /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(6702..6806)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      961 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1021 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1081 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1141 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1201 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1261 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1321 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1381 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1441 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1501 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1561 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgaat
     1621 gctccaaaac tcagcagtgc ttctggtgct ggtgatcagt gcttctgcaa cccatgaggc
     1681 ggagcagaat gactctgtga gccccaggaa atcccgagtg gcggctcaaa actcagctga
     1741 agtggttcgt tgcctcaaca gtgctctaca ggtcggctgc ggggcttttg catgcctgga
     1801 aaactccacc tgtgacacag atgggatgta tgacatctgt aaatccttct tgtacagcgc
     1861 tgctaaattt gacactcagg gaaaagcatt cgtcaaagag agcttaaaat gcatcgccaa
     1921 cggggtcacc tccaaggtct tcctcgccat tcggaggtgc tccactttcc aaaggatgat
     1981 tgctgaggtg caggaagagt gctacagcaa gctgaatgtg tgcagcatcg ccaagcggaa
     2041 ccctgaagcc atcactgagg tcgtccagct gcccaatcac ttctccaaca gatactataa
     2101 cagacttgtc cgaagcctgc tggaatgtga tgaagacaca gtcagcacaa tcagagacag
     2161 cctgatggag aaaattgggc ctaacatggc cagcctcttc cacatcctgc agacagacca
     2221 ctgtgcccaa acacacccac gagctgactt caacaggaga cgcaccaatg agccgcagaa
     2281 gctgaaagtc ctcctcagga acctccgagg tgaggaggac tctccctccc acatcaaacg
     2341 cacatcccat gagagtgcat aagaattctg cagtcgacgg taccgcgggc ccgggatcct
     2401 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2461 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2521 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2581 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2641 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2701 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3121 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3481 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3541 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3601 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3661 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3721 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3781 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3841 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3901 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3961 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4021 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4081 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4141 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4201 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4261 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4321 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4381 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4441 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4501 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4621 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     5641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6841 ggatcgggag atctcccgat cccctatggt
//
