LOCUS       V044983                 7038 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044983
VERSION     V044983
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..1718
                     /label="Styxl1(NM_029659)"
                     /note="Styxl1(NM_029659)"
                     /gene="Styxl1"
     CDS             1725..1790
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1814..2402
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2468..2516
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2718..3146
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3160..3489
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3556..4350
                     /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    4526..4647
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4696..4712)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4720..4736
                     /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(4744..4774)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4789..4810
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5098..5686)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5857..6717)
                     /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(6718..6822)
                     /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 tgacctgctt ttctgcgagc
      781 caacagaact ctataacatc ttgaatcagg tctccaagct gtccagattg gctgagccca
      841 actacctctg tttactggat gtccgatcaa aacgacagta tgacgagagc catgtgatca
      901 ctgccctcag agtgaagaag cgagatcatc agtacctcat cccggagtcc gtagacctgg
      961 agtgtgtgag atactgcatc gtgtacgaca gcaacaccag ttccctggag ctgtgcatac
     1021 ggcggcagct ggaggaggag gaagaggagg aggaggtaga aagagaggac aaggaggagg
     1081 acactgaact cctacctgga cctgctgtgg aatttggaca aatcctcatt cactttaccc
     1141 gccaacctgt ctatgtcctc agagggggct atgaatgctt ctctggcatg taccacttct
     1201 tccggactca gaagatcatc tggatgcccc aggaactgga tgccttccag ccatacccag
     1261 ttgagatact tccaggcaga gtctacctgg gcaaaatcag tcaagcctgc aatgctaaga
     1321 tgcacaagga cttgaaaatt aaagcacatg tcaatatttc catggagaca acaccctatt
     1381 ttataggcaa tgcagacaag ctcttacata tcaagcttga ggattctcca gacaccttgc
     1441 tttttgacta tttccgccac atctgtcact ttatagaact tcacctcagg ctccactctg
     1501 tcgttctggt cttttccacc cggggcatca gccgcagcgt tgctgctgtg gtggccttac
     1561 tcatgcatta taatgaggag acgctgaaga ggtcctgggc ccaagtgaag aaatgcaaaa
     1621 ccaacatgcg tccgaatcgg gccttcgtgg ctcagctgct ggagtgggag aaagtcctcc
     1681 atggggaata tctcacagac atctccgagc ctatctacgg atccgactac aaagaccatg
     1741 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     1801 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1861 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1921 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     1981 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2041 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2101 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2161 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2221 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2281 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2341 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2401 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2461 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2521 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2581 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2641 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     2701 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3061 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3241 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3421 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3481 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3541 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3601 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3661 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3721 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3781 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3841 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3901 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3961 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4021 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4081 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4141 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4201 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4261 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4321 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4381 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4441 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4501 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4561 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4621 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4681 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4741 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4801 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4861 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4921 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4981 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5101 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5161 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5221 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5281 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5341 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5401 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5461 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5521 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5581 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5641 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5701 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5761 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5821 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5881 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5941 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6001 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6061 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6121 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6181 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6241 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6301 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6361 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6421 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6481 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6541 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6601 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6661 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6721 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6781 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6841 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     6901 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6961 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7021 atgaagaatc tgcttagg
//
