LOCUS       V044241                 8961 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044241
VERSION     V044241
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..3641
                     /label="Sart3(NM_016926)"
                     /note="Sart3(NM_016926)"
                     /gene="Sart3"
     CDS             3648..3713
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3737..4325
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4391..4439
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4641..5069
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5083..5412
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5479..6273
                     /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    6449..6570
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6619..6635)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6643..6659
                     /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(6667..6697)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6712..6733
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7021..7609)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7780..8640)
                     /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(8641..8745)
                     /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 ccaccatggc gacgacggcc gcatcttcgg
      781 cgtccgagcc ggaggttgag ccccaggccg ggcctgaggc cgagggagaa gaggatgagg
      841 cgaagccggc cggtgtgcag cggaaggtgc tgtccggcgc tgtggccgcg gaggcggcgg
      901 aggccaaggg ccccggatgg gacctgcagc gggaaggcgc gagcgggagc gatggggatg
      961 aggaggacgc catggcttcc tccgccgaga gctccgccgg ggaggacgag tgggagtacg
     1021 acgaggagga ggagaagaac cagctggaga tcgagcggct ggaggagcag ctgtccatca
     1081 atggctatga ttacaactgc cacgtggagc tcatcaggct gctgcggctg gaaggcgagc
     1141 tcagcagagt gagggcggcc cgccagaaga tgagtgagct cttccccctg accgaagagc
     1201 tctggctgga gtggctccac gatgagatca gcatggccat ggacggcctg gaccgcgagc
     1261 acgtgtacga gctctttgag agagccgtga aggactacat ctgtccaaac atttggctag
     1321 agtatggcca gtactcagtt ggtggcattg gtcagaaagg tggccttgag aaggttcgct
     1381 ctgtctttga aagagccctg tcctctgttg gcctgcacat gacgaaaggc ctggccatct
     1441 gggaggccta ccgagagttt gaaagcgcca tcgtggaggc tgctcggctg gagaaagtgc
     1501 acagtctctt ccggcgacag ctggcgatcc cactgtacga gatggaggcc acctttgcag
     1561 agtatgaaga atggtcagag gagcccatgc cggagtctgt acttcagagc tatcagaaag
     1621 cgctggggca gctagagaag tacaagcctt acgaggaagc gctgctgcag gcagaggccc
     1681 ctcggctggc ggaataccaa gcttacatcg acttcgagat gaaaatcggg gatcctgccc
     1741 gtattcagtt gatctttgag cgtgccctgg tggagaactg cctggttcca gacttatgga
     1801 tccgctacag tcagtaccta gatcgacagc tgaaagttaa ggacttggtt ttatctgtac
     1861 acagccgtgc tgtgcggaac tgcccatgga cagttgccct gtggagtcgg taccttctgg
     1921 ccatggagcg acatggactg gaccatcaaa cgatttctgc gaccttcgag aacgctctga
     1981 gtgccggctt catccaggcc actgactatg tggagatctg gcaggtgtac ctcgactacc
     2041 tgaggagaag ggttgacttc agacaggact ctagcaagga gctggaagag ctgcggtcca
     2101 tgttcacgcg agctctggag tacctgcagc aggaggttga ggagcgtttc agcgagagtg
     2161 gggatccaag ctgcctgatc atgcagagct gggctcgggt tgaggctcgc ctgtgcaata
     2221 acatgcagaa agcccgagag ctctgggaca gcatcatgac cagagggaat gccaagtacg
     2281 ccaacatgtg gctggagtat tacaacctgg aacgggcaca cggtgacaca caacactgtc
     2341 ggaaggctct gcaccgagct gtccagtgca cgagtgacta ccctgagcac gtctgtgaag
     2401 tgttgctcac catggagagg acagaaggga ccttagaaga ttgggatcta gccattcaga
     2461 aaacggagac gcgcttggct cgtgtgaatg agcagagaat gaaggccgca gagaaggaag
     2521 cagctcttgt gcagcaggaa gaagaaaagg ccgagcagcg gaagaaggtg cgggcggaga
     2581 agaaagccct gaaaaagaag aagaaaacgc gaggtgccga caagcgcagg gaggacgagg
     2641 acgaggagaa cgagtggggc gaagaggagg aagagcagcc ttccaaacgc agaaggacgg
     2701 agaacagtct ggcctctgga gaggcttcgg ctatgaagga agaaacagag ctctccggga
     2761 aatgcttaac gatagatgtt ggtcctcctt ccaagcagaa agagaaggca gcctccctta
     2821 agcgggacat gcccaaggtg gctcacgaca gcagtaagga cagtgtcacc gtgtttgtca
     2881 gcaacctgcc ctacagcata gaagagcccg aggtgaagct caggccgctc tttgaggtct
     2941 gtggggaggt ggtccagatc aggccaattt tcagcaaccg cggggacttc cggggctact
     3001 gctatgtgga gtttggagag gagaagtcag cccagcaggc cctggagctg gacaggaaga
     3061 ttgtggaggg caggccgatg tttgtgtccc cctgtgtgga taagagcaaa aaccctgatt
     3121 ttaaggtgtt cagatacagt accaccctgg agaaacacaa actcttcatc tctggcctgc
     3181 ccttttcctg caccaaagag gagctcgagg acatttgtaa ggcccacggc accgtcaagg
     3241 acctcaggct ggtcactaac agggctggca agccgaaggg cctggcgtat gtggagtatg
     3301 aaaacgagtc ccaggcgtcc caggcagtga tgaagatgga cggcatgacc atcagagaga
     3361 atgtcatcaa ggtggcaatc agcaatcccc ctcagcgaaa agtcccagag aagccagaag
     3421 tgaggacagc accaggggcc cccatgctcc cccggcagat gtatggcgcg cgcgggaagg
     3481 gacggaccca gctctctctt cttcctcgag ctctgcagcg ccagggtgct gctcctcagg
     3541 ctgagaacgg cccagctccg gggcccgcgg tcgccccgtc tgtggccaca gaggctccta
     3601 agatgtccaa tgctgatttt gcgaagttgc ttctgagaaa gggatccgac tacaaagacc
     3661 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     3721 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3781 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3841 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3901 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3961 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4021 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4081 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     4141 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4201 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4261 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4321 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4381 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4441 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4501 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4561 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     4621 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4681 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4741 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     4801 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4861 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4921 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4981 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     5041 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5101 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5161 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5221 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5281 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5341 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5401 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5461 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5521 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5581 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5641 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5701 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5761 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5821 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5881 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5941 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6001 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6061 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6121 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6181 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6241 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6301 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6361 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6421 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6481 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6541 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6601 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7201 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8761 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     8821 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     8881 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     8941 tgcatgaaga atctgcttag g
//
