LOCUS       V036836                 6984 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036836
VERSION     V036836
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..1664
                     /label="MTCH2(NM_014342)"
                     /note="MTCH2(NM_014342)"
                     /gene="MTCH2"
     CDS             1671..1736
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1760..2348
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2414..2462
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2664..3092
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3106..3435
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3502..4296
                     /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    4472..4593
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4642..4658)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4666..4682
                     /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(4690..4720)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4735..4756
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5044..5632)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5803..6663)
                     /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(6664..6768)
                     /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 ggacgcggcc agtcaggtgc
      781 tcctgggctc cggtctcacc atcctgtccc agccgctcat gtacgtgaaa gtgctcatcc
      841 aggtgggata tgagcctctt cctccaacaa taggacgaaa tatttttggg cggcaagtgt
      901 gtcagcttcc tggtctcttt agttatgctc agcacattgc cagtatcgat gggaggcgcg
      961 ggttgttcac aggcttaact ccaagactgt gttcgggagt ccttggaact gtggtccatg
     1021 gtaaagtttt acagcattac caggagagtg acaagggtga ggagttagga cctggaaatg
     1081 tacagaaaga agtctcatct tcctttgacc acgttatcaa ggagacaact cgagagatga
     1141 tcgctcgttc tgctgctacc ctcatcacac atcccttcca tgtgatcact ctgagatcta
     1201 tggtacagtt cattggcaga gaatccaagt actgtggact ttgtgattcc ataataacca
     1261 tctatcggga agagggcatt ctaggatttt tcgcgggtct tgttcctcgc cttctaggtg
     1321 acatcctttc tttgtggctg tgtaactcac tggcctacct cgtcaatacc tatgcactgg
     1381 acagtggggt ttctaccatg aatgaaatga agagttattc tcaagctgtc acaggatttt
     1441 ttgcgagtat gttgacctat ccctttgtgc ttgtctccaa tcttatggct gtcaacaact
     1501 gtggtcttgc tggtggatgc cctccttact ccccaatata tacgtcttgg atagactgtt
     1561 ggtgcatgct acaaaaagag gggaatatga gccgaggaaa tagcttattt ttccggaagg
     1621 tcccctttgg gaagacttat tgttgtgacc tgaaaatgtt aattggatcc gactacaaag
     1681 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     1741 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1801 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1861 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1921 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1981 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2041 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2101 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2161 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2221 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2281 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2341 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2401 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2461 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     2521 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2581 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     2641 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2701 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2761 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2821 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2881 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2941 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3001 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3061 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3181 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3301 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3361 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3421 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3481 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3541 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3601 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3661 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3721 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3781 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3841 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3901 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3961 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4021 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4081 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4141 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4201 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4261 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4321 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4381 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4441 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4501 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4561 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4621 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4681 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4741 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4801 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4861 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4921 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4981 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5041 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5101 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5161 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5221 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5281 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5341 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5401 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5461 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5521 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5581 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5641 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5701 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5761 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5821 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5881 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5941 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6001 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6061 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6121 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6181 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6241 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6301 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6361 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6421 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6481 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6541 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6601 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6661 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6721 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6781 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     6841 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     6901 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     6961 aattgcatga agaatctgct tagg
//
