LOCUS       V047653                 7653 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047653
VERSION     V047653
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             706..2379
                     /label="GPC1(NM_002081)"
                     /note="GPC1(NM_002081)"
                     /gene="GPC1"
     CDS             2386..2409
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2429..3017
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3083..3131
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3333..3761
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3775..4104
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4171..4965
                     /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    5141..5262
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5311..5327)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5335..5351
                     /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(5359..5389)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5404..5425
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5713..6301)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6472..7332)
                     /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(7333..7437)
                     /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 gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga gctccgggcc
      721 cgaggctggt ggctgctatg tgcggccgca gcgctggtcg cctgcgcccg cggggacccg
      781 gccagcaaga gccggagctg cggcgaggtc cgccagatct acggagccaa gggcttcagc
      841 ctgagcgacg tgccccaggc ggagatctcg ggtgagcacc tgcggatctg tccccagggc
      901 tacacctgct gcaccagcga gatggaggag aacctggcca accgcagcca tgccgagctg
      961 gagaccgcgc tccgggacag cagccgcgtc ctgcaggcca tgcttgccac ccagctgcgc
     1021 agcttcgatg accacttcca gcacctgctg aacgactcgg agcggacgct gcaggccacc
     1081 ttccccggcg ccttcggaga gctgtacacg cagaacgcga gggccttccg ggacctgtac
     1141 tcagagctgc gcctgtacta ccgcggtgcc aacctgcacc tggaggagac gctggccgag
     1201 ttctgggccc gcctgctcga gcgcctcttc aagcagctgc acccccagct gctgctgcct
     1261 gatgactacc tggactgcct gggcaagcag gccgaggcgc tgcggccctt cggggaggcc
     1321 ccgagagagc tgcgcctgcg ggccacccgt gccttcgtgg ctgctcgctc ctttgtgcag
     1381 ggcctgggcg tggccagcga cgtggtccgg aaagtggctc aggtccccct gggcccggag
     1441 tgctcgagag ctgtcatgaa gctggtctac tgtgctcact gcctgggagt ccccggcgcc
     1501 aggccctgcc ctgactattg ccgaaatgtg ctcaagggct gccttgccaa ccaggccgac
     1561 ctggacgccg agtggaggaa cctcctggac tccatggtgc tcatcaccga caagttctgg
     1621 ggtacatcgg gtgtggagag tgtcatcggc agcgtgcaca cgtggctggc ggaggccatc
     1681 aacgccctcc aggacaacag ggacacgctc acggccaagg tcatccaggg ctgcgggaac
     1741 cccaaggtca acccccaggg ccccgggcct gaggagaagc ggcgccgggg caagctggcc
     1801 ccgcgggaga ggccaccttc aggcacgctg gagaagctgg tctccgaagc caaggcccag
     1861 ctccgcgacg tccaggactt ctggatcagc ctcccaggga cactgtgcag tgagaagatg
     1921 gccctgagca ctgccagtga tgaccgctgc tggaacggga tggccagagg ccggtacctc
     1981 cccgaggtca tgggtgacgg cctggccaac cagatcaaca accccgaggt ggaggtggac
     2041 atcaccaagc cggacatgac catccggcag cagatcatgc agctgaagat catgaccaac
     2101 cggctgcgca gcgcctacaa cggcaacgac gtggacttcc aggacgccag tgacgacggc
     2161 agcggctcgg gcagcggtga tggctgtctg gatgacctct gcagccggaa ggtcagcagg
     2221 aagagctcca gctcccggac gcccttgacc catgccctcc caggcctgtc agagcaggaa
     2281 ggacagaaga cctcggctgc cagctgcccc cagcccccga ccttcctcct gcccctcctc
     2341 ctcttcctgg cccttacagt agccaggccc cggtggcggc tcgagcacca tcaccatcac
     2401 catcaccatt agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2461 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2521 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2581 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2641 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2701 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2761 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2821 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2881 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2941 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3001 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3061 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3121 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3181 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3241 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     3301 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3481 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3661 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4021 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4081 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4141 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4321 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4681 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4981 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5041 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5101 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5281 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5341 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5401 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5461 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5521 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5881 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6121 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7441 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7501 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7561 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7621 ttgaccgaca attgcatgaa gaatctgctt agg
//
