LOCUS       V045142                 7479 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045142
VERSION     V045142
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..2159
                     /label="Dmap1(NM_023178)"
                     /note="Dmap1(NM_023178)"
                     /gene="Dmap1"
     CDS             2166..2231
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2255..2843
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2909..2957
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3159..3587
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3601..3930
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3997..4791
                     /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    4967..5088
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5137..5153)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5161..5177
                     /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(5185..5215)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5230..5251
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5539..6127)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6298..7158)
                     /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(7159..7263)
                     /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 tacgggcgca gatgtacgag
      781 acattctaga actcgggggt ccagagggag atgccgcctc tgggaccatc agcaaaaagg
      841 atattatcaa cccggacaag aaaaagtcca agaagtcctc agagacgctg accttcaaga
      901 ggcctgaggg catgcatcgg gaggtctatg ctttgcttta ctctgacaaa aaggatgcac
      961 ccccactgct gcccagtgac actggtcagg ggtatcggac agtgaaggcg aaactggggt
     1021 ccaagaaggt tcgcccttgg aaatggatgc cttttactaa cccagctcga aaggacggcg
     1081 ctatgttttt ccactggcga cgagcggcgg aggagggcaa ggactaccct tttgccaggt
     1141 tcaataagac ggtgcaggtg cccgtgtact cagagcagga gtaccaactc taccttcatg
     1201 atgacgcatg gactaaggca gagactgacc acctatttga cctcagccgc cgatttgatc
     1261 tgcgcttcgt agttattcac gatcggtatg accaccagca gttcaagaag cgttctgtgg
     1321 aggacctgaa agagaggtac taccacattt gtgccaagct tgccaacgtg agggctgtgc
     1381 caggcacaga tctcaagata ccagtgtttg atgctgggca tgagagacgg cggaaggaac
     1441 agctagagcg gctttacaac cgaaccccag agcaggtggc agaggaggag tacctcctac
     1501 aggagctgcg taagattgag gcccggaaaa aagagcggga gaagcgcagc caagacctgc
     1561 agaagctgat tacagcagca gacaccactg cagagcagcg gcgcacggaa cgcaaggctc
     1621 ccaagaagaa gctaccccaa aagaaggagg ctgagaagcc ggctgtccct gagactgcag
     1681 gcatcaagtt tccagatttt aagtcggcag gtgtcacgct acggagccag cggatgaagc
     1741 tacccagctc tgtgggtcag aagaagatca aggcgctgga acagatgctg ctggaacttg
     1801 gtgtggagct gagccctacc cccacagagg agctggtgca tatgttcaat gagttgcgga
     1861 gcgacctggt gttactctac gagctcaagc aggcctgtgc caactgtgaa tatgagctac
     1921 agatgctgcg gcaccggcac gaggccctgg ctcgggcagg agtgctgggg gcccctgccg
     1981 cagcagcagt gggaccaacc ccggcttctg ctgagccaac agtgtctgaa tctggacttg
     2041 gtctggaccc caccaaggat accatcattg atgtcgtggg tgcacccctc acacccaatt
     2101 cgcggaaacg acgggaatca gcctccagct catcttctgt gaagaaagcc aagaaaccag
     2161 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     2221 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2281 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2341 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2401 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2461 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2521 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2581 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2641 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2701 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2761 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2821 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     2881 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2941 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3001 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3061 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3121 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3181 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3241 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3301 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3361 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3421 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3481 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3541 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3601 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3661 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3721 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3781 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3841 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3901 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3961 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4021 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4081 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4141 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4201 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4261 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4321 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4381 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4441 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4501 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4561 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4621 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4681 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4741 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4801 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4861 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4921 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4981 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5041 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5101 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5161 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5221 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5281 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5701 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5941 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7261 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7321 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7381 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7441 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
