LOCUS       V045909                 7191 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045909
VERSION     V045909
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..1871
                     /label="Mthfsd(NM_001166482)"
                     /note="Mthfsd(NM_001166482)"
                     /gene="Mthfsd"
     CDS             1878..1943
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1967..2555
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2621..2669
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2871..3299
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3313..3642
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3709..4503
                     /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    4679..4800
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4849..4865)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4873..4889
                     /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(4897..4927)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4942..4963
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5251..5839)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6010..6870)
                     /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(6871..6975)
                     /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 gacccaggag ggtgtctcca
      781 aacaaagcat acgtgaacgg atttgggact acatggaatc ccatgatata gctgactttc
      841 cccgacccgt tcatcacagg atccccaact tcaagggtgc tgctcaggct gcggggcacc
      901 tcccacacct gcaagctttc catgtggcca gaaccattaa agtcaacccg gacgctcctc
      961 agagaaacgc ccgcttcctg gtgctggaga gcaaaaaaac actgctggtc ccaacaccac
     1021 gactgcgaac agggctgttt aacaagatca cgccaccccc tggagccacc aaagacattc
     1081 tgaggaaatg tgccacctcc cagggtgtgc ggaatttcag cgtcccagtg ggcttggatt
     1141 ccagtgttct tgtggatcta gttgtggtgg gatctgttgc tgtgtctgag aaaggctggc
     1201 gaattgggaa aggggaaggc tatgctgatc ttgaatacgc catgatggtg tcaatgggag
     1261 ccgtccacaa ggggacaccg gttgtcacaa ttgtccacga ctgccaggtc gtagacatcc
     1321 ccgaggctct tgtggaggac cacgacctca cagttgacta catcctcact cccaccaggg
     1381 tcatcactac aggctgtgca cgcccaaagc caacgggcat catgtggtcc aaggtcagct
     1441 gtgagatgct tacgaaaatc ccagtgctca ggaacctccg ggagagagag aaacaggctg
     1501 ggaaggatgt cactctccgg gacgagcctg gcagccagca gccagctcca ggccccatca
     1561 ggagacccca ggatagacca caaacaggtt ctcgaggagg gtcccgtagt cccctgcagg
     1621 gggcggacac ccaactagct gccactgtgt gtgtgggaaa cctgcccttc actactcgtg
     1681 tgcgtgagct gaagagagtc ctccaggagc tgggggtggt gcccctgagg cttacctggc
     1741 aggggccaca gcacagggca gttctacact atacagactc agctgctgcc cagcaggccg
     1801 cctctctcct acagggccta cgtctgggtg ccaacgcctt gagggtgtca ctggggcagc
     1861 agagggatat gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     1921 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     1981 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2041 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2101 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2161 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2221 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2281 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2341 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2401 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2461 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2521 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2581 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2641 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2701 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2761 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2821 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     2881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2941 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3001 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3061 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3121 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3181 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3241 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3301 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3481 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3541 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3601 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3661 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3721 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3781 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3841 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3901 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3961 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4021 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4081 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4141 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4201 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4261 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4321 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4381 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4441 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4501 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4561 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4621 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4681 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4741 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4801 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5821 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5881 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5941 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6001 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6061 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6961 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7021 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7081 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7141 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
