LOCUS       V022001                 9919 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022001
VERSION     V022001
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             1666..5406
                     /label="NID1(NM_002508)"
                     /note="NID1(NM_002508)"
                     /gene="NID1"
     polyA_signal    5486..5710
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5756..6184
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6198..6527
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6594..7388
                     /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    7562..7683
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7732..7748)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7756..7772
                     /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(7780..7810)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7825..7846
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8134..8719)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8890..9750)
                     /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(9751..9855)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagcttg gtaccatgtt ggcctcgagc
     1681 agccggatcc gggctgcgtg gacgcgggcg ctgctgctgc cgctgctgct ggcggggcct
     1741 gtgggctgcc tgagccgcca ggagctcttt cccttcggcc ccggacaggg ggacctggag
     1801 ctggaggacg gggatgactt cgtctctcct gccctggagc tgagtggggc gctccgcttc
     1861 tacgacagat ccgacatcga cgcagtctac gtcaccacaa atggcatcat tgctacgagt
     1921 gaacccccgg ccaaagaatc ccatcccggg ctcttcccac caacattcgg tgcagtcgcc
     1981 cctttcctgg cggacttgga cacgaccgat ggcctgggga aggtttatta tcgagaagac
     2041 ttatccccct ccatcactca gcgagcagca gagtgtgtcc acagagggtt cccggagatc
     2101 tctttccagc ctagtagcgc ggtggttgtc acttgggaat ccgtggcccc ctaccaaggg
     2161 cccagcaggg acccagacca gaaaggcaag agaaacacgt tccaggctgt tctagcctcc
     2221 tctgattcca gctcctatgc cattttcctt tatcctgagg atggtctgca gttccatacg
     2281 acattctcaa agaaggaaaa caaccaagtt cctgccgtgg ttgcattcag tcaaggttca
     2341 gtgggattct tatggaagag caacggagct tataacatat ttgctaatga cagggaatca
     2401 gttgaaaatt tggccaagag tagtaactct gggcagcagg gtgtctgggt gtttgagatt
     2461 gggagtccag ccaccaccaa tggcgtggtg cctgcagacg tgatcctcgg aactgaagat
     2521 ggggcagagt atgatgatga ggatgaagat tatgacctgg cgaccactcg tctgggcctg
     2581 gaggatgtgg gcaccacgcc cttctcctac aaggctctga gaaggggagg tgctgacaca
     2641 tacagtgtgc ccagcgtcct ctccccgcgc cgggcagcta ccgaaaggcc ccttggacct
     2701 cccacagaga gaaccaggtc tttccagttg gcagtggaga cttttcacca gcagcaccct
     2761 caggtcatag atgtggatga agttgaggaa acaggagttg ttttcagcta taacacggat
     2821 tcccgccaga cgtgtgctaa caacagacac cagtgctcgg tgcacgcaga gtgcagggac
     2881 tacgccacgg gcttctgctg cagctgtgtc gctggctata cgggcaatgg caggcaatgt
     2941 gttgcagaag gttcccccca gcgagtcaat ggcaaggtga aaggaaggat ctttgtgggg
     3001 agcagccagg tccccattgt ctttgagaac actgacctcc actcttacgt agtaatgaac
     3061 cacgggcgct cctacacagc catcagcacc attcccgaga ccgttggata ttctctgctt
     3121 ccactggccc cagttggagg catcattgga tggatgtttg cagtggagca ggacggattc
     3181 aagaatgggt tcagcatcac cgggggtgag ttcactcgcc aggctgaggt gaccttcgtg
     3241 gggcacccgg gcaatctggt cattaagcag cggttcagcg gcatcgatga gcatgggcac
     3301 ctgaccatcg acacggagct ggagggccgc gtgccgcaga ttccgttcgg ctcctccgtg
     3361 cacattgagc cctacacgga gctgtaccac tactccacct cagtgatcac ttcctcctcc
     3421 acccgggagt acacggtgac tgagcccgag cgagatgggg catctccttc acgcatctac
     3481 acttaccagt ggcgccagac catcaccttc caggaatgcg tccacgatga ctcccggcca
     3541 gccctgccca gcacccagca gctctcggtg gacagcgtgt tcgtcctgta caaccaggag
     3601 gagaagatct tgcgctatgc tctcagcaac tccattgggc ctgtgaggga aggctcccct
     3661 gatgctcttc agaatccctg ctacatcggc actcatgggt gtgacaccaa cgcggcctgt
     3721 cgccctggtc ccaggacaca gttcacctgc gagtgctcca tcggcttccg aggagacggg
     3781 cgaacctgct atgatattga tgaatgttca gaacaaccct cagtgtgtgg gagccacaca
     3841 atctgcaata atcacccagg aaccttccgc tgcgagtgtg tggagggcta ccagttttca
     3901 gatgagggaa cgtgtgtggc tgtcgtggac cagcgcccca tcaactactg tgaaactggc
     3961 cttcataact gcgacatacc ccagcgggcc cagtgtatct acacaggagg ctcctcctac
     4021 acctgttcct gcttgccagg cttttctggg gatggccaag cctgccaaga tgtagatgaa
     4081 tgccagccaa gccgatgtca ccctgacgcc ttctgctaca acactccagg ctctttcacg
     4141 tgccagtgca aacctggtta tcagggagac ggcttccgtt gcgtgcccgg agaggtggag
     4201 aaaacccggt gccagcacga gcgagaacac attctcgggg cagcgggggc gacagaccca
     4261 cagcgaccca ttcctccggg gctgttcgtt cctgagtgcg atgcgcacgg gcactacgcg
     4321 cccacccagt gccacggcag caccggctac tgctggtgcg tggatcgcga cggccgcgag
     4381 gtggagggca ccaggaccag gcccgggatg acgcccccgt gtctgagtac agtggctccc
     4441 ccgattcacc aaggacctgc ggtgcctacc gccgtgatcc ccttgcctcc tgggacccat
     4501 ttactctttg cccagactgg gaagattgag cgcctgcccc tggagggaaa taccatgagg
     4561 aagacagaag caaaggcgtt ccttcatgtc ccggctaaag tcatcattgg actggccttt
     4621 gactgcgtgg acaagatggt ttactggacg gacatcactg agccttccat tgggagagct
     4681 agtctacatg gtggagagcc aaccaccatc attagacaag atcttggaag tccagaaggt
     4741 atcgctgttg atcaccttgg ccgcaacatc ttctggacag actctaacct ggatcgaata
     4801 gaagtggcga agctggacgg cacgcagcgc cgggtgctct ttgagactga cttggtgaat
     4861 cccagaggca ttgtaacgga ttccgtgaga gggaaccttt actggacaga ctggaacaga
     4921 gataacccca agattgaaac ttcctacatg gacggcacga accggaggat ccttgtgcag
     4981 gatgacctgg gcttgcccaa tggactgacc ttcgatgcgt tctcatctca gctctgctgg
     5041 gtggatgcag gcaccaatcg ggcggaatgc ctgaacccca gtcagcccag cagacgcaag
     5101 gctctcgaag ggctccagta tccttttgct gtgacgagct acgggaagaa tctgtatttc
     5161 acagactgga agatgaattc cgtggttgct ctcgatcttg caatttccaa ggagacggat
     5221 gctttccaac cccacaagca gacccggctg tatggcatca ccacggccct gtctcagtgt
     5281 ccgcaaggcc ataactactg ctcagtgaac aatggcggct gcacccacct atgcttggcc
     5341 accccaggga gcaggacctg ccgttgccct gacaacacct tgggagttga ctgtatcgaa
     5401 cagaaatgag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     5461 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5521 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5581 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5641 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5701 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     5761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6181 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6541 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6601 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6661 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6721 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6781 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6841 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6901 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6961 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7021 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7081 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7141 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7201 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7261 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7321 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7381 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     7441 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7501 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7561 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7621 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7681 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7741 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7801 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     8341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8641 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     8701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     9001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     9061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     9121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9901 tctcccgatc ccctatggt
//
