LOCUS       V021990                 9698 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021990
VERSION     V021990
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             683..4354
                     /label="Copa(NM_009938)"
                     /note="Copa(NM_009938)"
                     /gene="Copa"
     CDS             4361..4450
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4474..5062
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5128..5176
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5378..5806
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5820..6149
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6216..7010
                     /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    7186..7307
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7356..7372)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7380..7396
                     /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(7404..7434)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7449..7470
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7758..8346)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8517..9377)
                     /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(9378..9482)
                     /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 ggatccgcca ccatgctaac caaattcgag acgaagagcg cgcgggtcaa
      721 aggccttagt tttcatccta aaagaccctg gatcctgacc agtttacaca atggggtcat
      781 ccagttatgg gactatcgga tgtgcaccct aattgacaaa tttgatgaac atgatggtcc
      841 agtgcgtggc attgacttcc ataagcagca gccactgttc gtctctggag gcgatgatta
      901 taagattaag gtttggaatt acaaacttcg gcgctgcctg ttcacattgc tcgggcactt
      961 ggattatatc cgtacaacgt ttttccatca tgaatatcct tggattctga gcgcctcaga
     1021 tgatcagacc attcgggtgt ggaactggca gtctagaacc tgtgtctgtg tgttaacggg
     1081 gcacaaccat tatgtaatgt gtgctcagtt ccacccctct gaagacctgg tggtatcagc
     1141 tagcctggac cagactgtcc gcgtttggga tatttctggt ctaaggaaga aaaacttgtc
     1201 tcctggtgcg gtggagtcgg atgtgagagg aataactggc gttgatctgt ttggaaccac
     1261 agatgccgtg gtgaagcatg tgctagaggg tcatgaccgt ggagtcaact gggctgcctt
     1321 ccacccgact atgcccctta ttgtatctgg ggcagatgac cggcaagtga agatctggcg
     1381 tatgaatgaa tcaaaggcat gggaggttga cacctgccgg ggccattaca acaatgtgtc
     1441 ttgtgctgtt tttcaccctc gccaagagtt gatcctcagc aattctgaag acaagagtat
     1501 tcgagtctgg gatatgtcta agcggacggg ggttcagact ttccgtaggg atcatgatcg
     1561 cttctgggtc ctggctgccc atcccaacct taacctcttt gcagcaggcc atgatggtgg
     1621 gatgattgta tttaagctgg aacgtgagcg tccagcctat gctgttcatg gtaacatgct
     1681 gcattatgta aaggaccgat tcctccgtca gctggatttc aacagctcca aagatgtagc
     1741 tgtgatgcag ttgcggagcg gttctaagtt tccagtattc aacatgtcat acaatccagc
     1801 agaaaacgca gtgctgctct gcaccagagc aagcaatcta gagaacagta cctacgacct
     1861 atacaccatc cccaaagacg cagactccca gaaccctgac gcccctgaag ggaaacgatc
     1921 ctcaggcctg acagcggttt gggttgctcg gaatcggttt gctgtcctag atcgcatgca
     1981 ttcacttctg attaaaaacc tgaaaaatga gatcaccaag aaaatccagg tgccaaactg
     2041 tgatgaaatc ttctatgctg gtacaggcaa cctcctgctt cgggatgcag attccatcac
     2101 actctttgac gtccagcaga agcgaactct ggcatcagta aagatttcca aggtgaaata
     2161 tgttatctgg tcggcagaca tgtctcatgt agcgttactg gccaagcatg ccattgtgat
     2221 ctgtaaccgc aagctggatg ctctgtgtaa cattcatgag aacattcgtg tcaagagtgg
     2281 ggcctgggat gaaagtgggg tgtttatcta taccacaagc aaccacatca aatatgctgt
     2341 caccactggg gaccatggga ttatccgaac tctggattta cccatctatg tcaccagagt
     2401 gaagggcaat aatgtatact gcctagacag agagtgtcgt cctcgggtgc taaccattga
     2461 tcccactgag ttcaagttca aactggccct gatcaacaga aagtatgatg aggtactcca
     2521 catggtgagg aacgctaagc tggtaggcca gtcaatcatc gcttatctcc agaagaaggg
     2581 ctatcctgaa gtggcgctgc attttgtaaa ggatgagaaa actcgtttca gcctggccct
     2641 ggagtgtgga aacattgaga ttgctctaga ggcagccaaa gccctggacg acaagaactg
     2701 ctgggagaag ctaggagaag tggccctgct acagggcaat caccaaattg tagagatgtg
     2761 ctatcagcgc accaaaaact ttgacaaact ttccttcctg taccttatta ctggcaacct
     2821 ggagaaactt cgaaagatga tgaagatcgc tgagatccga aaggacatga gtggccatta
     2881 tcagaatgcc ctatacctag gtgatgtgtc agagagagta cggatcctga agaactgtgg
     2941 gcagaaatcc ctagcctatc tcacagctgc tacccatggc ttagatgaag aagccgagag
     3001 cctgaaggaa acatttgacc ctgagaagga gacaatccca gacattgatc ctaatgccaa
     3061 gctgctccag ccacctgcac ctatcatgcc attggataca aactggccgt tactgaccgt
     3121 gtccaaaggg ttctttgaag gctccattgc aagcaagggg aagggaggag cattggctgc
     3181 tgacattgac atcgacactg taggcactga aggctgggga gaggacgcag agctgcagct
     3241 ggatgaagat gggtttgtgg aggctccaga aggtttggga gaggatgttc ttggaaaggg
     3301 acaagaagaa ggaggtggct gggatgtaga agaagatttg gagctcccac ctgagctgga
     3361 tgtaccctct ggggtgtctg gtagtgctga agatgggttc tttgtgcccc caaccaaggg
     3421 aacaagccca acgcaaatct ggtgcaataa ctcccagctt ccagtagatc acatcttggc
     3481 aggttctttt gaaacagcca tgcggcttct ccatgatcag gtgggagtaa tccagtttgg
     3541 cccctataag caactgttcc tacagacata tgcccgaggc cgcaccacct atcaagcgct
     3601 gccctgcttg ccctccatgt atagttaccc taatcgcaat tggaaggatg cagggctaaa
     3661 gaatggagta ccagctgtag gcctaaagct taatgacctg atccaacgtt tgcagctgtg
     3721 ttatcagctc accacagttg gcaagtttga ggaagctgtg gaaaaattcc gttccatcct
     3781 actcagtgtg cctcttcttg ttgtggacaa taaacaggag attgcagagg cccagcagct
     3841 catcaccatt tgccgtgaat acattgtggg tttgtgcatg gagatagaaa ggaagaagct
     3901 acctaaggag accctagacc agcagaagcg catctgtgag atggcagcct acttcacaca
     3961 ctcaaatctg cagcctgtgc atatgatctt ggtgctgcgg acagccctca accttttctt
     4021 caagctcaag aacttcaaga cagccgccac ctttgcccga cgcctgctgg aacttgggcc
     4081 caagcctgaa gtagctcagc agacaagaaa aatcttatct gcctgtgaga aaaaccccac
     4141 agatgcctgc cagctcaatt atgacatgca caaccccttt gacatttgtg ctgcatctta
     4201 ccggcccatc taccgtggaa aaccagtgga gaagtgtcca ctcagtgggg cctgctattc
     4261 ccctgagttc aaggggcaaa tctgcagggt cactacagtg acagagattg gcaaagatgt
     4321 gattgggctg aggatcagtc ctcttcagtt tcgcctcgag gaacaaaaac taatatcaga
     4381 ggaagatttg gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga
     4441 ggaggactta tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     4501 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4561 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4621 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4681 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4741 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4801 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4861 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4921 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4981 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     5041 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     5101 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     5161 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5221 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5281 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5341 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5521 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     5761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5821 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5881 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5941 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6061 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     6121 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     6181 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6241 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6301 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6361 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     6421 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6481 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6541 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6601 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6661 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6721 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6781 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6841 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6901 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6961 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     7021 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     7081 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     7141 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     7201 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7261 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     7321 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7381 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7441 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7501 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7561 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7621 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7681 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7741 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7801 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7861 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7921 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7981 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8041 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8101 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8161 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     8221 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8281 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8341 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8401 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8461 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8521 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8581 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8641 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8701 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8761 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8821 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8881 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8941 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9001 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9061 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9121 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9181 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9241 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9301 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9361 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9421 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9481 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     9541 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     9601 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     9661 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
