LOCUS       V041960                 6583 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041960
VERSION     V041960
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             744..770
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             774..800
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             804..830
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             837..1256
                     /label="Pfn1(NM_011072)"
                     /note="Pfn1(NM_011072)"
                     /gene="Pfn1"
     misc_feature    1359..1947
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2013..2061
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2263..2691
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2705..3034
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3101..3895
                     /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    4071..4192
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4241..4257)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4265..4281
                     /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(4289..4319)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4334..4355
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5402..6262)
                     /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(6263..6367)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgccacc atgtacccat acgacgtacc tgattatgca acttatcctt
      781 atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca aagcttatgg
      841 ccgggtggaa cgcctacatc gacagcctta tggcggacgg gacctgtcag gacgcggcca
      901 tcgtaggcta caaggactcg ccctccgtct gggccgccgt ccccgggaag accttcgtta
      961 gcattacgcc agctgaggtt ggtgtcctgg taggcaaaga ccggtcaagt tttttcgtca
     1021 atgggctgac acttgggggc cagaaatgtt ctgtgatccg ggactcactg ctgcaagacg
     1081 gggaatttac aatggatctt cgtaccaaga gcaccggagg agcccccacc ttcaatgtca
     1141 ctgtcaccat gactgccaag acgctagtcc tgctgatggg caaagaaggt gtccacggtg
     1201 gtttgatcaa caagaaatgt tatgaaatgg cctctcacct gcggcgttcc cagtactgag
     1261 aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc cttaagggtt
     1321 cgatccctac cggttagtaa tgagtttgat atctcgacaa tcaacctctg gattacaaaa
     1381 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     1441 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     1501 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     1561 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     1621 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     1681 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1741 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     1801 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     1861 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     1921 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     1981 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     2041 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     2101 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     2161 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     2221 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2401 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     2461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     2641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2941 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3001 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3061 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3121 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3181 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3241 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3301 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3361 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3421 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3481 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3541 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3601 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3661 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3721 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3781 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3841 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3901 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3961 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4021 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4081 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4141 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4201 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4261 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4321 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4381 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4441 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4501 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4561 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6361 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6421 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6481 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6541 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
