LOCUS       V022885                10571 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022885
VERSION     V022885
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             1597..6231
                     /label="ABCC2(NM_000392)"
                     /note="ABCC2(NM_000392)"
                     /gene="ABCC2"
     polyA_signal    6333..6557
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6603..7031
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7045..7374
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7441..8040
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    8214..8335
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8384..8400)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8408..8424
                     /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(8432..8462)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8477..8498
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8786..9371)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9542..10402)
                     /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(10403..10507)
                     /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 aagcttatgc tggagaagtt ctgcaactct
     1621 actttttgga attcctcatt cctggacagt ccggaggcag acctgccact ttgttttgag
     1681 caaactgttc tggtgtggat tcccttgggc tacctatggc tcctggcccc ctggcagctt
     1741 ctccacgtgt ataaatccag gaccaagaga tcctctacca ccaaactcta tcttgctaag
     1801 caggtattcg ttggttttct tcttattcta gcagccatag agctggccct tgtactcaca
     1861 gaagactctg gacaagccac agtccctgct gttcgatata ccaatccaag cctctaccta
     1921 ggcacatggc tcctggtttt gctgatccaa tacagcagac aatggtgtgt acagaaaaac
     1981 tcctggttcc tgtccctatt ctggattctc tcgatactct gtggcacttt ccaatttcag
     2041 actctgatcc ggacactctt acagggtgac aattctaatc tagcctactc ctgcctgttc
     2101 ttcatctcct acggattcca gatcctgatc ctgatctttt cagcattttc agaaaataat
     2161 gagtcatcaa ataatccatc atccatagct tcattcctga gtagcattac ctacagctgg
     2221 tatgacagca tcattctgaa aggctacaag cgtcctctga cactcgagga tgtctgggaa
     2281 gttgatgaag agatgaaaac caagacatta gtgagcaagt ttgaaacgca catgaagaga
     2341 gagctgcaga aagccaggcg ggcactccag agacggcagg agaagagctc ccagcagaac
     2401 tctggagcca ggctgcctgg cttgaacaag aatcagagtc aaagccaaga tgcccttgtc
     2461 ctggaagatg ttgaaaagaa aaaaaagaag tctgggacca aaaaagatgt tccaaaatcc
     2521 tggttgatga aggctctgtt caaaactttc tacatggtgc tcctgaaatc attcctactg
     2581 aagctagtga atgacatctt cacgtttgtg agtcctcagc tgctgaaatt gctgatctcc
     2641 tttgcaagtg accgtgacac atatttgtgg attggatatc tctgtgcaat cctcttattc
     2701 actgcggctc tcattcagtc tttctgcctt cagtgttatt tccaactgtg cttcaagctg
     2761 ggtgtaaaag tacggacagc tatcatggct tctgtatata agaaggcatt gaccctatcc
     2821 aacttggcca ggaaggagta caccgttgga gaaacagtga acctgatgtc tgtggatgcc
     2881 cagaagctca tggatgtgac caacttcatg cacatgctgt ggtcaagtgt tctacagatt
     2941 gtcttatcta tcttcttcct atggagagag ttgggaccct cagtcttagc aggtgttggg
     3001 gtgatggtgc ttgtaatccc aattaatgcg atactgtcca ccaagagtaa gaccattcag
     3061 gtcaaaaata tgaagaataa agacaaacgt ttaaagatca tgaatgagat tcttagtgga
     3121 atcaagatcc tgaaatattt tgcctgggaa ccttcattca gagaccaagt acaaaacctc
     3181 cggaagaaag agctcaagaa cctgctggcc tttagtcaac tacagtgtgt agtaatattc
     3241 gtcttccagt taactccagt cctggtatct gtggtcacat tttctgttta tgtcctggtg
     3301 gatagcaaca atattttgga tgcacaaaag gccttcacct ccattaccct cttcaatatc
     3361 ctgcgctttc ccctgagcat gcttcccatg atgatctcct ccatgctcca ggccagtgtt
     3421 tccacagagc ggctagagaa gtacttggga ggggatgact tggacacatc tgccattcga
     3481 catgactgca attttgacaa agccatgcag ttttctgagg cctcctttac ctgggaacat
     3541 gattcggaag ccacagtccg agatgtgaac ctggacatta tggcaggcca acttgtggct
     3601 gtgataggcc ctgtcggctc tgggaaatcc tccttgatat cagccatgct gggagaaatg
     3661 gaaaatgtcc acgggcacat caccatcaag ggcaccactg cctatgtccc acagcagtcc
     3721 tggattcaga atggcaccat aaaggacaac atcctttttg gaacagagtt taatgaaaag
     3781 aggtaccagc aagtactgga ggcctgtgct ctcctcccag acttggaaat gctgcctgga
     3841 ggagatttgg ctgagattgg agagaagggt ataaatctta gtgggggtca gaagcagcgg
     3901 atcagcctgg ccagagctac ctaccaaaat ttagacatct atcttctaga tgaccccctg
     3961 tctgcagtgg atgctcatgt aggaaaacat atttttaata aggtcttggg ccccaatggc
     4021 ctgttgaaag gcaagactcg actcttggtt acacatagca tgcactttct tcctcaagtg
     4081 gatgagattg tagttctggg gaatggaaca attgtagaga aaggatccta cagtgctctc
     4141 ctggccaaaa aaggagagtt tgctaagaat ctgaagacat ttctaagaca tacaggccct
     4201 gaagaggaag ccacagtcca tgatggcagt gaagaagaag acgatgacta tgggctgata
     4261 tccagtgtgg aagagatccc cgaagatgca gcctccataa ccatgagaag agagaacagc
     4321 tttcgtcgaa cacttagccg cagttctagg tccaatggca ggcatctgaa gtccctgaga
     4381 aactccttga aaactcggaa tgtgaatagc ctgaaggaag acgaagaact agtgaaagga
     4441 caaaaactaa ttaagaagga attcatagaa actggaaagg tgaagttctc catctacctg
     4501 gagtacctac aagcaatagg attgttttcg atattcttca tcatccttgc gtttgtgatg
     4561 aattctgtgg cttttattgg atccaacctc tggctcagtg cttggaccag tgactctaaa
     4621 atcttcaata gcaccgacta tccagcatct cagagggaca tgagagttgg agtctacgga
     4681 gctctgggat tagcccaagg tatatttgtg ttcatagcac atttctggag tgcctttggt
     4741 ttcgtccatg catcaaatat cttgcacaag caactgctga acaatatcct tcgagcacct
     4801 atgagatttt ttgacacaac acccacaggc cggattgtga acaggtttgc cggcgatatt
     4861 tccacagtgg atgacaccct gcctcagtcc ttgcgcagct ggattacatg cttcctgggg
     4921 ataatcagca cccttgtcat gatctgcatg gccactcctg tcttcaccat catcgtcatt
     4981 cctcttggca ttatttatgt atctgttcag atgttttatg tgtctacctc ccgccagctg
     5041 aggcgtctgg actctgtcac caggtcccca atctactctc acttcagcga gaccgtatca
     5101 ggtttgccag ttatccgtgc ctttgagcac cagcagcgat ttctgaaaca caatgaggtg
     5161 aggattgaca ccaaccagaa atgtgtcttt tcctggatca cctccaacag gtggcttgca
     5221 attcgcccgg agctggttgg gaacctgact gtcttctttt cagccttgat gatggttatt
     5281 tatagagata ccctaagtgg ggacactgtt ggctttgttc tgtccaatgc actcaatatc
     5341 acacaaaccc tgaactggct ggtgaggatg acatcagaaa tagagaccaa cattgtggct
     5401 gttgagcgaa taactgagta cacaaaagtg gaaaatgagg caccctgggt gactgataag
     5461 aggcctccgc cagattggcc cagcaaaggc aagatccagt ttaacaacta ccaagtgcgg
     5521 taccgacctg agctggatct ggtcctcaga gggatcactt gtgacatcgg tagcatggag
     5581 aagattggtg tggtgggcag gacaggagct ggaaagtcat ccctcacaaa ctgcctcttc
     5641 agaatcttag aggctgccgg tggtcagatt atcattgatg gagtagatat tgcttccatt
     5701 gggctccacg acctccgaga gaagctgacc atcatccccc aggaccccat cctgttctct
     5761 ggaagcctga ggatgaatct cgaccctttc aacaactact cagatgagga gatttggaag
     5821 gccttggagc tggctcacct caagtctttt gtggccagcc tgcaacttgg gttatcccac
     5881 gaagtgacag aggctggtgg caacctgagc ataggccaga ggcagctgct gtgcctgggc
     5941 agggctctgc ttcggaaatc caagatcctg gtcctggatg aggccactgc tgcggtggat
     6001 ctagagacag acaacctcat tcagacgacc atccaaaacg agttcgccca ctgcacagtg
     6061 atcaccatcg cccacaggct gcacaccatc atggacagtg acaaggtaat ggtcctagac
     6121 aacgggaaga ttatagagtg cggcagccct gaagaactgc tacaaatccc tggacccttt
     6181 tactttatgg ctaaggaagc tggcattgag aatgtgaaca gcacaaaatt ctaggatcca
     6241 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     6301 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     6361 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6421 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6481 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     6541 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     6601 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6661 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6721 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6781 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6841 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6901 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6961 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     7021 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     7081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     7141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7201 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7261 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     7321 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     7381 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     7441 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     7501 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     7561 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     7621 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     7681 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7741 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7801 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     7861 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7921 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7981 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     8041 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     8101 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     8161 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     8221 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8281 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8341 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     8401 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8461 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8521 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8581 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8641 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8701 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8761 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8821 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8881 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8941 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     9001 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9121 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9181 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     9241 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9301 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     9361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     9421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     9481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    10021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    10081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    10141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    10201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    10261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    10321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    10381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    10441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    10501 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
    10561 tcccctatgg t
//
