LOCUS       V022676                 8581 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022676
VERSION     V022676
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             649..1365
                     /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             1387..3324
                     /label="Cep72(NM_028959)"
                     /note="Cep72(NM_028959)"
                     /gene="Cep72"
     misc_feature    3357..3945
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4011..4059
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4261..4689
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4703..5032
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5099..5893
                     /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    6069..6190
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6239..6255)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6263..6279
                     /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(6287..6317)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6332..6353
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6641..7229)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7400..8260)
                     /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(8261..8365)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccatgg cgccaggcca gcggttggtt ttgtgtgaag agactgtacg agagcgcagc
     1441 ggcctgggtc cccaccgcga cctggctgaa cttcgatctt tatctattcc tggaacatac
     1501 caagagaaga ttactcacct gggaaactct ctgatgcact tgacagctct aaaatcctta
     1561 gacctctcac gtaactcctt ggttagcctt gagggtatac agtatctggt ctccttggag
     1621 agtctcaatc tctactacaa ctgtatttct tcattagcag aagtatttag gctccacacc
     1681 ctactggagc tccaggacgt ggacttccgg cttaaccccg ttgtgaagaa tgagtctgac
     1741 taccgcctgt ttgtcgtgca catgctcccg aagctccgac agctagatga ccgtcctgtg
     1801 agagaaagtg agcggaaagc ttcccagctt cattttgccc cagaagattc actgaactca
     1861 aaggagaact tctcaaccac tttaaccgtt ggaagaccac accacctcag aaacagatgt
     1921 actgagacct cggccaaaaa atgcttggtc atggatgctg atgatgaggc tgtcctgaac
     1981 cttattgccg agtgtgaatg ggacctaagc aatcctcctg ggaacatgag ctccagccaa
     2041 aaggaacatg aagctgacct ccattatgcc caagagtcca gacatctgtt gagtcctttg
     2101 tccatccagc accagtgtgg ggattcagcg aggcgaggcc atgagaagaa gaaagtcacc
     2161 tccagagggt gtccaggtca ttcaccgcag gaccagctct gtggggaact ccccctacag
     2221 catggcttgc cagaggcctg ccatatgcac gtacagcatg cccgcatcac ttcacagcca
     2281 gactctgtgg atgttgagga ctgtgcttca tctgctcaga aatcaagttt gtcatcacaa
     2341 aaaacagtaa atcctttgcc tgtccctgaa aagtacagga agcgaaggat gcctggtggg
     2401 agattccagg tgccttcaga ccgggagtgt ttgagcttcc tagagagggc tgacgggccc
     2461 tccagccttg aggacagtct aagcagacag gatggcttgg agggccagag tcaggtggcc
     2521 ttgtctcact ctgaggcttt ggaggctgaa gagcggacat ctcatggcac aagtgacccc
     2581 agagtgctgt ctcctaaact ctgctcggca gcggtgcctg agcggaggtc cactctggaa
     2641 gtggcccttc tggaggcgct tctcgacctg atagacaggt gctcaagtgg cagtggatcc
     2701 ctgcatggca atgaggcctt ccttgctcag gccaagcatg ttttgtcatc tcttcaagag
     2761 ttcacagcaa ctcgggacag ttcagcactt gaaaaggaag gaattggtta cctatctctt
     2821 gaaaataaga ctctgcaaag ccgccttgct gagcaacagc aacagtatac tgctacagtg
     2881 accaagatga cagcagagct caataataca aagagggagc tggacacatt gagacaacat
     2941 ttggacaaat cattggaaga gaatagccac ttaaagtctc ttctgtacaa tgtgaaaaaa
     3001 gaagtaaaga ctgcagacac ttcaactgca ctcactttgc agatcactgg acttcaggca
     3061 agtatgaagc agctctctgg ggaagttgtg gagctgaagc aacacgtgga gcactatgac
     3121 aagatccagg agctcacaca gatgcttcaa gagagccaca gctccctggt aagcaccaat
     3181 gagcatctcc tgcaggagct gagccgcaca cgggcacagc atagagctga ggtagagcag
     3241 atgcgctgga gcttccagga atttaagaag actacggccc tgatccctca ccgtagctcc
     3301 cgacgtgggg gccggcagtc ctgctagctc gagtctagag ggcccgatat ctcgacaatc
     3361 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3421 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3481 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3541 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3601 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3661 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3721 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     3781 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3841 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3901 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     3961 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4021 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4081 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4141 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4201 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     4261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4321 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4381 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     4441 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4561 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4621 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4681 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4801 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4861 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4921 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4981 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     5041 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     5101 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5161 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5221 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5281 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5341 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5401 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5461 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5521 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5581 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5641 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5701 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5761 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5821 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5881 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5941 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6001 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6061 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6181 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7081 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8401 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     8461 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8521 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8581 g
//
