LOCUS       V030655                 8437 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030655
VERSION     V030655
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..3096
                     /label="AFG3L2(NM_006796)"
                     /note="AFG3L2(NM_006796)"
                     /gene="AFG3L2"
     CDS             3103..3129
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3133..3159
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3163..3189
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3213..3801
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3867..3915
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4117..4545
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4559..4888
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4955..5749
                     /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    5925..6046
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6095..6111)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6119..6135
                     /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(6143..6173)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6188..6209
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6497..7085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7256..8116)
                     /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(8117..8221)
                     /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 ccaccatggc gcaccgctgt
      721 ttgcggctgt ggggccgggg cggctgctgg ccccgcggcc tacagcagct cctcgtgcct
      781 ggcggcgtgg gcccgggcga gcagccctgc ctccggacgc tttaccgatt tgttacaact
      841 caagcaaggg ccagcagaaa ttctcttttg acagatataa ttgctgctta tcaaagattc
      901 tgttctcgac ccccaaaagg atttgaaaaa tactttccta atggaaaaaa tggaaaaaaa
      961 gctagtgaac ctaaagaagt tatgggagag aaaaaagaat caaagccagc tgctaccaca
     1021 cgctcttctg gaggaggagg tggtggcggt ggaaaacgag gtggcaagaa agatgattct
     1081 cactggtggt ccaggtttca gaagggtgac attccatggg acgacaagga tttcaggatg
     1141 ttcttcctct ggactgctct gttctggggt ggagtcatgt tttacttgct gctcaagaga
     1201 tccgggagag aaatcacttg gaaggacttt gtcaataact atctttcaaa aggagtagta
     1261 gacagattgg aagtcgtcaa caagcgtttt gttcgagtga cctttacacc aggaaaaact
     1321 cctgttgatg ggcaatacgt ttggtttaat attggcagtg tggacacctt tgaacggaat
     1381 ctggaaactt tacagcagga attgggcata gaaggagaaa atcgggtgcc tgttgtctac
     1441 attgctgaaa gtgatggctc ttttctgctg agcatgctgc ctacggtgct catcatcgcc
     1501 ttcttgctct acaccatcag aagagggcct gctggcattg gccggacagg ccgagggatg
     1561 ggcggactct tcagtgtcgg agaaaccact gccaaggtct taaaggatga aattgatgtg
     1621 aagttcaaag atgtggctgg ctgtgaggag gccaagctag agatcatgga atttgtgaat
     1681 ttcttgaaaa acccaaagca gtatcaagac ctaggagcaa aaatcccaaa gggtgccatt
     1741 ctcactggtc ctccaggcac tgggaagacg ctgctagcta aggccacagc cggagaagcc
     1801 aatgtcccct tcatcaccgt tagtggatct gagtttttgg agatgttcgt tggtgtgggc
     1861 cctgctagag tccgagactt atttgccctt gctcggaaga atgccccttg catcctcttc
     1921 atcgatgaaa tcgatgcggt gggaaggaag agaggaagag gcaactttgg agggcagagt
     1981 gagcaggaga acacactcaa ccagctgctg gtggagatgg atggttttaa tacaacaaca
     2041 aatgtcgtca ttttggccgg caccaatcga ccagatatcc tggaccccgc gctgcttagg
     2101 ccggggcgtt tcgacaggca gatctttatt ggaccaccag acataaaagg aagagcttct
     2161 attttcaaag ttcatctccg accgctaaaa ctggacagta ccctggagaa ggataaattg
     2221 gcaagaaaac tggcatcttt aactccaggg ttttcaggtg ctgatgttgc taatgtctgt
     2281 aatgaagctg cgttgattgc tgcaaggcat ctgtcagatt ccataaatca gaaacacttt
     2341 gaacaggcaa ttgaacgagt gattggtggc ttagagaaga aaacgcaggt tctgcagcct
     2401 gaggagaaga agactgtggc ataccacgaa gcaggccatg cggttgccgg ctggtatctg
     2461 gagcacgcag acccgctttt aaaggtatcc atcatcccac gtggcaaagg actaggttat
     2521 gctcagtatt taccaaaaga acaatacctc tataccaaag agcagctctt ggataggatg
     2581 tgtatgactt taggtggtcg agtctctgaa gaaatcttct ttggaagaat tacaactggt
     2641 gctcaagatg acttgagaaa agtaactcag agtgcatatg cccaaattgt tcagtttggc
     2701 atgaatgaaa aggttgggca aatctccttt gacctcccac gtcaggggga catggtattg
     2761 gagaaacctt acagtgaagc cactgcaaga ttgatagatg atgaagtacg aatacttatt
     2821 aatgatgctt ataaaagaac agtagctctt ctcacagaaa agaaagctga cgtggagaag
     2881 gttgctcttc tgttgttaga aaaagaagta ttagataaga atgatatggt tgaacttttg
     2941 ggccccagac catttgcgga aaaatctacc tatgaagaat ttgtggaagg cactggcagc
     3001 ttggatgagg acacctcact tccagaaggc cttaaggact ggaacaagga gcgggaaaag
     3061 gagaaagagg agcccccggg tgagaaagtt gccaacctcg agtacccata cgacgtacct
     3121 gattatgcaa cttatcctta tgatgtacct gattatgcta gctatcctta tgatgtccca
     3181 gactacgcat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     3241 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3301 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3361 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3421 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3481 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3541 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3601 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3661 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3721 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3781 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     3841 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     3901 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3961 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4021 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4081 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4141 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4201 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4261 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4321 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4381 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4441 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     4501 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4561 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4681 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4741 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4801 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4861 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4921 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4981 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5041 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5101 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5161 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5221 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5281 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5341 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5401 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5461 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5521 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5581 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5641 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5701 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5761 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5821 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5881 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5941 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6001 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6061 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6661 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6901 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8221 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     8281 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     8341 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8401 aggcttgacc gacaattgca tgaagaatct gcttagg
//
