LOCUS       V020050                 7160 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020050
VERSION     V020050
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..2653
                     /label="MTDH(NM_178812)"
                     /note="MTDH(NM_178812)"
                     /gene="MTDH"
     CDS             2663..2680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2727..2951
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2997..3425
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3439..3768
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3835..4629
                     /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    4803..4924
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4973..4989)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4997..5013
                     /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(5021..5051)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5066..5087
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5375..5960)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6131..6991)
                     /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(6992..7096)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gctgcacgga gctggcagga cgagctggcc cagcaggccg aggagggctc
      961 ggcccggctg cgggaaatgc tctcggtcgg cctaggcttt ctgcgcaccg agctgggcct
     1021 cgacctgggg ctggagccga aacggtaccc cggctgggtg atcctggtgg gcactggcgc
     1081 gctcgggctg ctgctgctgt ttctgctggg ctacggctgg gccgcggctt gcgccggcgc
     1141 ccgcaaaaag cggaggagcc cgccccgcaa gcgggaggag gcggcggccg tgccggccgc
     1201 ggcccccgac gacctggcct tgctgaagaa tctccggagc gaggaacaga agaagaagaa
     1261 ccggaagaaa ctgtccgaga agcccaaacc aaatgggcgg actgttgaag tggctgaggg
     1321 tgaagctgtt cgaacacctc aaagtgtaac agcaaagcag ccaccagaga ttgacaagaa
     1381 aaatgaaaag tcaaagaaaa ataagaagaa atcaaagtca gatgctaaag cagtgcaaaa
     1441 cagttcacgc catgatggaa aggaagttga tgaaggagcc tgggaaacta aaattagtca
     1501 cagagagaaa cgacagcagc gtaaacgtga taaggtgctg actgattctg gttcattgga
     1561 ttcaactatc cctgggatag aaaataccat cacagttacc accgagcaac ttacaaccgc
     1621 atcatttcct gttggttcca agaagaataa aggtgattct catctaaatg ttcaagttag
     1681 caactttaaa tctggaaaag gagattctac acttcaggtt tcttcaggat tgaatgaaaa
     1741 cctcactgtc aatggaggag gctggaatga aaagtctgta aaactctcct cacagatcag
     1801 tgcaggtgag gagaagtgga actccgtttc acctgcttct gcaggaaaga ggaaaactga
     1861 gccatctgcc tggagtcaag acactggaga tgctaataca aatggaaaag actggggaag
     1921 gagttggagt gaccgttcaa tattttctgg cattgggtct actgctgagc cagtttctca
     1981 gtctaccact tctgattatc agtgggatgt tagccgtaat caaccctata tcgatgatga
     2041 atggtctggg ttaaatggtc tgtcttctgc tgatcccaac tctgattgga atgcaccagc
     2101 agaagagtgg ggcaattggg tagacgaaga aagagcttca cttctaaagt cccaggaacc
     2161 aattcctgat gatcaaaagg tctcagatga tgataaagaa aagggagagg gagctcttcc
     2221 aactgggaaa tccaaaaaga aaaaaaagaa aaagaagaag caaggtgaag ataactctac
     2281 tgcacaggac acagaagaat tagaaaaaga gattagagaa gaccttccag tgaatacctc
     2341 taaaacccgt ccaaaacagg aaaaagcttt ttccttgaag accataagca ctagtgatcc
     2401 agccgaagta ctcgtcaaaa atagccagcc tatcaagact cttccacctg ctacttctac
     2461 cgagccatct gtaatcttat caaaaagtga ttctgacaag agctcttccc aagtgccgcc
     2521 aatactacaa gagacagata aatccaagtc aaataccaag caaaatagtg tgcctccttc
     2581 acagaccaag tctgaaacta gctgggaatc tcccaaacaa ataaaaaaga agaaaaaagc
     2641 cagacgagaa acggcggccg cacatcacca tcatcaccat tgactcgagt ctagagggcc
     2701 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2761 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2821 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2881 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2941 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3061 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3301 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3361 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3421 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3601 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3661 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3721 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3781 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4621 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4681 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4801 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4861 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4981 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5041 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5101 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5161 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     5941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7141 atctcccgat cccctatggt
//
