LOCUS       V042756                10158 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042756
VERSION     V042756
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..4838
                     /label="Spata31d1b(NM_001167593)"
                     /note="Spata31d1b(NM_001167593)"
                     /gene="Spata31d1b"
     CDS             4845..4910
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4934..5522
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5588..5636
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5838..6266
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6280..6609
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6676..7470
                     /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    7646..7767
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7816..7832)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7840..7856
                     /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(7864..7894)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7909..7930
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8218..8806)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8977..9837)
                     /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(9838..9942)
                     /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 ccaccatgga gaagattctc tcagctctga
      781 atggcctcag tgagacatgg ctgaccctag gctcagcttc ctaccacata gacctcaact
      841 acactcttct gagtggactg gggctactgc ttctgtacat ctcctacctg atattaaaac
      901 tgcttataca cttgctgtgg agagagaaat acaccccaaa gctatgggag aaagggaaga
      961 agagaaggca aaccctcaaa ggtgggcgag cttgccagag agaagcggcg cagaggaggg
     1021 agctgcagac aacccagcca agctctctgg aagagccctc tgaggcctcc tgcaatgaac
     1081 taccactcca cagagatccc ctcttgcctg tgtgtgatag atctggcaat atcagtcatc
     1141 agctctccca ggaccacctc aaagatggta aagcctctta cttgtgcgtg gcttccacag
     1201 ttcctgggac ggaggcctcg tttgttctgt cctctctgct cacaggaaac caggcagggc
     1261 attcggctcc agcccttcta ccggagcctt taccgctggc tccgtccatc ctttcaccta
     1321 actgctttgc ccccctagaa ggcttctact cagccacact gctgcacgac tctctgacac
     1381 cagagtccac acttcccttg aacccttcac ttctcccgga ccccacccca ccctatccac
     1441 ttgcctcttt tcctcttcca acaagagaag tgcagaagtc agaagtggtc ttccaactgg
     1501 aaaccatgag atctctggtg gacagactca atgagatgcc cactaatgtc ccaccagccc
     1561 aaggcactgg ctgtgtaaga caaacaatgc cagaggcccc ccaacagcag cctgctgctg
     1621 acaaccaatt atcttcagac tcaaaaaacc agtatctccc agctcctcat gttcctcaag
     1681 tccctttgca gggagacaca agggcctgcc atgtagagcc tggatacctc ttttttccca
     1741 actctaatgc cgtgaaactg ttccaaaggc tagacaagaa agtgagtgat ttcctgatgg
     1801 tgaaggggga gcaggaggag tgggaggagg aagaggacaa aggatgtttt ataaagtcac
     1861 ccagttcttc ggagaagctg ctatcaggag agacatcggg tgagccacag gattttgtgg
     1921 cttctcattc gtcaagcagc aaaggcaagc tcacgcacca gccgctccct cactccaaga
     1981 cttttgaaga ccatgaggag cttcagtctt ccctgctctt ctgggggccc ccctctctac
     2041 acagtgaggc cctgagccct actaccacta acgcctatga ccgttcctca gcattggtct
     2101 gcttcaacag catggctgag gcctccattg ctgactactc tcatggctgt atgcttccca
     2161 cacctttgca tgcatatcaa ccacagacat ggctacagac cgaaccccag tcccatcccc
     2221 aacgtgaccc tcctgccgag ttgcagcctc agcctcaacc ccaatcccgg gtctcaatct
     2281 tgacattttc tcatcaatcc gagcctcgag actgtggggt gcattttcac agaccccagg
     2341 gagacgagca gcttctcagt tcttctgaga tgcagtgctt ggaatacaac atcttgaaaa
     2401 aggaacagga gagggtctgg ggtttacccc tcatggtcaa aaagtccaag tatacttttt
     2461 gtccctcacc tcccaaggtc ttgttgggaa gtcgctcctc caaggcctgt actcccaggc
     2521 ccatccttcc tggagacttc cccctcacca gtgagctcca gaagaaactt gaacaccatc
     2581 tccgaaagag actcatccaa caccgctggg gcctgcccga caggatcgac aagtccctgt
     2641 cactcatgag tccccagtca gaactgatcg acttctctga gtcaaaaaag aaccggggac
     2701 tctcttggat cactttcttt aagtaccaag gcaacaaaga ctctcaggtc ttatctggag
     2761 gtagctctcg aaatcggatg cctagaaggc attctctacg ggagactagg gtcaaagaac
     2821 gaacatatag tcaagatagg ggccacaatg accaatcctg gggtaacaac tttcaggagg
     2881 cttcacaaaa cagtgtgcag tctcacttga agacacaccg gaagagtcgt atgggcaggc
     2941 agcctcccag gttttcaagc ccctcacagg tgaaacagta ccagagagga attgaaaaag
     3001 ccctggaaaa acacctgaac aagaaaatta gggaaatcag caggggtgag atccctacca
     3061 cagtggatag gtcgaggcat tctgtgaaca tggcaaacat agcgcagcct ccaccagaga
     3121 cctcgcccaa acgaatgaaa gatctggcac caatggcatg taaggagcac atgctgacga
     3181 ggcaccccca cagcttagcg cttagtccta gcaaagagaa gatgctggaa gaacacatca
     3241 ccaccttccg cagaaggatg gcatttggcc tcccacaaag ggttgaggag tccttagagt
     3301 cttactcgac aaaagcagag ccttctcagc ctttccctca gctccacgtt caagcccaca
     3361 acgtttctag agcagaccct gccaagtcct ccaggtccct ccggagaaca actactggag
     3421 atagaatggg gacaatgact tcagtcccca cccaacagag gcctcttcct gctgcctcgc
     3481 tggtgggcca cagccagcca gcctctgaga acaagaaggt gggtgtagat agagacctta
     3541 gcgtaacccc aaggggcaga gagccaaccc agcactggac acccaggagg gcagacaaag
     3601 gtgacgtgca gcagtctgga tcacacaaca gacctggccc agagctgccc atgagcccag
     3661 gtgggccaac atatgagaga ctggcttcca gcaccaacac acaggagtct caggaagaaa
     3721 ggaggagccg ggaagacggc tccgaggctg agggctccac agagctacac aagggagagc
     3781 aactcccggg tcttcaccca cagtccacta agagcttgaa aggcacccaa ggcctgtgct
     3841 ccccagggag ccctgtgact gcatgtcagt ccccgcaggg aatgtcattt ctccacaact
     3901 ctgagagccc tgactctaaa agtcaggtgt ccacagaagt tcagctgaac tcagagggcg
     3961 ggacacacaa ccaagtgcca gacctgcctg ccacgccctt tgccccacaa gaaatgactt
     4021 ccaaacccca gggcccctcc ggtggggaca tggcagtttc tcaggtgctg cgtgtgcaca
     4081 tgcccactgt gggcatcagc atggagtcga ggcagggtcc ctgggtccct gcatatgtct
     4141 caggcaagag caagaacaag gactgtctcc ctgctgccag gggattgcca cagctggcaa
     4201 ctgaggcagg aaagtttggg ggaggggatg caggcttagg gacatcccag accacaggga
     4261 agggacactg tgttcaggcc agggcaccag aggagacaca aggactcaca gcctccccag
     4321 cactgacacc taagagtcag cctcaggaaa atcagttcac cagccaggtg aagggcttct
     4381 ggcagaggct gtcccctggg aagaaacaca aaggacagga aaagtccctg gccaagggct
     4441 gctctccatt agcatctggg aagggcacaa gccccatcaa agggagaagt gagttttgtg
     4501 ggaaccctga agctcagaac tgtgtgaggg agcctgggat ggtcctgagg aagcagctgg
     4561 ggcacaggca tgggacagtc accccctgcc cccaggtccc tgtgtctccc ctcatggggt
     4621 ctgaaggagc tcagcaggag gtgcccctgc aggctcaggc agagcctgtc cagaggctgc
     4681 cccacttgtg ctgcagaggt tcctgctctc cagtgccaag ggctgagtcc tgcagcccag
     4741 gacaggggca gacagcccct gagaggtgtg gccctacagg aaaagctaag agggtggaga
     4801 cctcccctgt gcttgcttct ccaccaaagt cctctctggg atccgactac aaagaccatg
     4861 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     4921 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4981 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5041 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5101 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5161 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5221 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5281 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     5341 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5401 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5461 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5521 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5581 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     5641 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5701 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5761 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     5821 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5881 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5941 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     6001 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6061 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6121 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6181 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     6241 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     6301 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6361 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6421 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6481 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6541 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6601 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     6661 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6721 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6781 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6841 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6901 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6961 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     7021 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     7081 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     7141 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     7201 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     7261 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     7321 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     7381 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7441 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     7501 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7561 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7621 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7681 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7741 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7801 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7861 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7921 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7981 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8041 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8101 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8161 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8221 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8281 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8341 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8401 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8461 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8521 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8581 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8641 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8701 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8761 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8821 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8881 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8941 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9001 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9061 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9121 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9181 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9241 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9301 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9361 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9421 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9481 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9541 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9601 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9661 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9721 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9781 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9841 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9901 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9961 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
    10021 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
    10081 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
    10141 atgaagaatc tgcttagg
//
