LOCUS       V041938                 8793 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041938
VERSION     V041938
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3473
                     /label="COPB2(NM_004766)"
                     /note="COPB2(NM_004766)"
                     /gene="COPB2"
     CDS             3480..3545
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3569..4157
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4223..4271
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4473..4901
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4915..5244
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5311..6105
                     /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    6281..6402
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6451..6467)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6475..6491
                     /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(6499..6529)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6544..6565
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6853..7441)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7612..8472)
                     /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(8473..8577)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc tctgcgactt gatatcaaaa
      781 gaaagctaac tgctagatct gatcgagtta agagtgtgga tctgcatcct acagagccat
      841 ggatgttggc aagtctttac aatggcagtg tgtgtgtttg gaatcatgaa acacagacac
      901 tggtgaagac atttgaagta tgtgatcttc ctgttcgagc tgcaaagttt gttgcaagga
      961 agaattgggt tgtgacagga gcggatgaca tgcagattag agtgttcaat tacaatactc
     1021 tggagagagt tcatatgttt gaagcacact cagactacat tcgctgtatt gctgttcatc
     1081 caacccagcc tttcattcta actagcagtg atgacatgct tattaagctc tgggactggg
     1141 ataaaaaatg gtcttgctca caagtgtttg aaggacacac ccattatgtt atgcagattg
     1201 tgatcaaccc caaagataac aatcagtttg ccagtgcctc tttggacagg actatcaagg
     1261 tgtggcagtt gggctcttcg tcaccaaact tcactttgga aggacatgag aaaggcgtga
     1321 attgcattga ttactacagt ggtggggaca agccatacct catttcaggt gcagatgacc
     1381 gtcttgttaa aatatgggat tatcagaata aaacatgtgt gcagacactg gaaggacatg
     1441 cccaaaatgt gtcttgtgcc agctttcatc ctgagttgcc aatcattatc acaggttcag
     1501 aagatggaac agtacgtatt tggcattcaa gcacctaccg gcttgagagc acactgaatt
     1561 atggaatgga gagggtatgg tgcgtggcca gtctaagagg gtcaaacaat gtcgctttgg
     1621 gctatgatga agggagcatc attgttaagc ttggtcggga ggaacctgcc atgtccatgg
     1681 atgccaatgg aaagataatt tgggccaagc attcagaagt ccagcaggcc aacctaaaag
     1741 caatgggaga tgctgaaatt aaagatggtg aaagattgcc actggcagta aaggatatgg
     1801 gcagttgtga aatataccct cagactattc agcacaatcc taatgggcgg tttgtggtgg
     1861 tgtgtggtga tggggagtat atcatctaca cagcaatggc attgagaaac aagagctttg
     1921 gatctgctca ggagtttgca tgggcccacg attcttcaga gtatgcaata agagagagca
     1981 acagcattgt aaagatattt aagaacttta aggaaaaaaa atcatttaaa ccagattttg
     2041 gagcagaaag tatctacggc ggcttcttat tgggagtcag atctgtaaat ggcttagcct
     2101 tctatgactg ggacaataca gaactcatac gaagaattga aattcagccc aaacatattt
     2161 tctggtctga ctctggagag ctagtctgta ttgctactga ggaatcattt tttatcctta
     2221 agtatctgtc agaaaaagtc ttggctgcac aggaaacaca tgagggagtt actgaagatg
     2281 gcattgaaga tgcctttgag gttcttggtg agattcagga aattgtgaaa acagggcttt
     2341 gggtaggcga ttgcttcatt tacacaagtt ctgtgaacag attaaattat tatgttggag
     2401 gagaaatagt caccattgcc cacttggaca ggacgatgta tctcctaggc tacattccta
     2461 aagacaacag gctttatctg ggggataaag aattgaacat cattagctat tccctgctgg
     2521 tttcagtcct ggaataccag acagctgtca tgcggaggga ctttagcatg gctgataagg
     2581 tccttcctac cattccaaaa gaacagagga ccagagttgc acactttttg gaaaagcagg
     2641 gcttcaagca gcaagctctt acagtatcca cagatcctga gcatcgtttt gagcttgctc
     2701 ttcagcttgg agagttaaaa attgcatacc agttagcagt ggaagcagag tcagaacaga
     2761 agtggaaaca acttgctgaa cttgccatta gtaaatgtca gtttggccta gcccaggagt
     2821 gcctgcatca tgcacaggat tatgggggcc tgctgctttt ggccactgcc tctggaaatg
     2881 ctaatatggt gaacaagcta gcagagggtg cggagagaga tggcaaaaat aatgtggcat
     2941 tcatgagcta ctttttacag ggcaaggttg atgcctgcct agagctctta attagaactg
     3001 gacggctgcc agaagctgcc ttcttggccc gaacttactt acccagtcag gtttcaaggg
     3061 tagtgaaact ctggagagag aatctctcaa aagtcaatca gaaagcagca gaatcccttg
     3121 ctgacccaac agagtatgaa aacctgttcc ctggattaaa agaagccttt gttgttgaag
     3181 aatgggtgaa ggaaacacat gctgatctgt ggccagccaa acaataccca cttgtcacgc
     3241 caaatgaaga gagaaatgtc atggaagagg gaaaagactt tcagccctca agatctacag
     3301 ctcaacagga acttgatggg aaacctgctt ctcctactcc ggttattgtg gcctcccaca
     3361 cagccaacaa agaagaaaag agtttactcg aactagaagt agatttggat aatttggaat
     3421 tagaagatat tgacacaaca gatatcaatc tggatgaaga tattttggat gatggatccg
     3481 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     3541 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3601 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3661 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3721 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3781 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3841 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3901 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3961 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4021 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4081 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4141 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     4201 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     4261 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4321 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4381 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     4441 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4501 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4561 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4621 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4681 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4741 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4801 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4861 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4921 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4981 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5041 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5101 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5161 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5221 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5281 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5341 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5401 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5461 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5521 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5581 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5641 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5701 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5761 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5821 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5881 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5941 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6001 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6061 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6121 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6181 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6241 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6301 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6361 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6421 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7021 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7261 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8581 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8641 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     8701 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8761 ttgaccgaca attgcatgaa gaatctgctt agg
//
