LOCUS       V044726                10486 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044726
VERSION     V044726
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"
     CDS             706..5142
                     /label="MRC2(NM_006039)"
                     /note="MRC2(NM_006039)"
                     /gene="MRC2"
     CDS             5149..5238
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    5262..5850
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5916..5964
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6166..6594
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6608..6937
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7004..7798
                     /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    7974..8095
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8144..8160)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8168..8184
                     /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(8192..8222)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8237..8258
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8546..9134)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9305..10165)
                     /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(10166..10270)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggg gcccggccgg
      721 ccggcccccg cgccctggcc tcgtcacctg ctgcgctgcg tcctgctcct cgggtgcctg
      781 cacctcggcc gtcccggcgc ccctggggac gccgccctcc cggaacccaa cgtcttcctc
      841 atcttcagcc atggactgca gggctgcctg gaggcccagg gcgggcaggt cagagtcacc
      901 ccggcttgca ataccagcct ccctgcccag cgctggaagt gggtctcccg aaaccggcta
      961 ttcaacctgg gtaccatgca gtgcctgggc acaggctggc caggcaccaa caccacggcc
     1021 tccctgggca tgtatgagtg tgaccgggaa gcactgaatc ttcgctggca ttgtcgtaca
     1081 ctgggtgacc agctgtcctt gctcctgggg gcccgcacca gcaacatatc caagcctggc
     1141 acccttgagc gtggtgacca gacccgcagt ggccagtggc gcatctacgg cagcgaggag
     1201 gacctatgtg ctctgcccta ccacgaggtc tacaccatcc agggaaactc ccacggaaag
     1261 ccgtgcacca tccccttcaa atatgacaac cagtggttcc acggctgcac cagcacgggc
     1321 cgcgaggatg gtcacctgtg gtgtgccacc acccaggact acggcaaaga cgagcgctgg
     1381 ggcttctgcc ccatcaagag taacgactgc gagaccttct gggacaagga ccagctgact
     1441 gacagctgct accagtttaa cttccagtcc acgctgtcgt ggagggaggc ctgggccagc
     1501 tgcgagcagc agggtgcgga tctgctgagc atcacggaga tccacgagca gacctacatc
     1561 aacggcctcc tcactgggta cagctccacc ctgtggatcg gcttgaatga cttggacacg
     1621 agcggaggct ggcagtggtc ggacaactcg cccctcaagt acctcaactg ggagagtgac
     1681 cagccggaca accccagtga ggagaactgt ggagtgatcc gcactgagtc ctcgggcggc
     1741 tggcagaacc gtgactgcag catcgcgctg ccctatgtgt gcaagaagaa gcccaacgcc
     1801 acggccgagc ccacccctcc agacaggtgg gccaatgtga aggtggagtg cgagccgagc
     1861 tggcagccct tccagggcca ctgctaccgc ctgcaggccg agaagcgcag ctggcaggag
     1921 tccaagaagg catgtctacg gggcggtggc gacctggtca gcatccacag catggcggag
     1981 ctggaattca tcaccaagca gatcaagcaa gaggtggagg agctgtggat cggcctcaac
     2041 gatttgaaac tgcagatgaa ttttgagtgg tctgacggga gccttgtgag cttcacccac
     2101 tggcacccct ttgagcccaa caacttccgg gacagtctgg aggactgtgt caccatctgg
     2161 ggcccggaag gccgctggaa cgacagtccc tgtaaccagt ccttgccatc catctgcaag
     2221 aaggcaggcc agctgagcca gggggccgcc gaggaggacc atggctgccg gaagggttgg
     2281 acgtggcaca gcccatcctg ctactggctg ggagaagacc aagtgaccta cagtgaggcc
     2341 cggcgcctgt gcactgacca tggctctcag ctggtcacca tcaccaacag gttcgagcag
     2401 gccttcgtca gcagcctcat ctacaactgg gagggcgagt acttctggac ggccctgcag
     2461 gacctcaaca gcaccggctc cttcttctgg ctcagtgggg atgaagtcat gtacacccac
     2521 tggaaccggg accagcccgg gtacagccgt gggggctgcg tggcgctggc cactggcagc
     2581 gccatggggc tgtgggaggt gaagaactgt acctcgttcc gggcccgcta catctgccgg
     2641 cagagcctgg gcactccagt gacgccggag ctgccggggc cagatcccac gcccagcctc
     2701 actggctcct gtccccaggg ctgggcctcg gacaccaaac tccggtattg ctataaggtg
     2761 ttcagctcag agcggctgca ggacaagaag agctgggtcc aggcccaggg ggcctgccag
     2821 gagctggggg cccagctgct gagcctggcc agctacgagg aggagcactt tgtggccaac
     2881 atgctcaaca agatcttcgg tgaatcagaa cccgagatcc acgagcagca ctggttctgg
     2941 atcggcctga accgtcggga tcccagaggg ggtcagagtt ggcgctggag cgacggcgta
     3001 gggttctctt accacaattt cgaccggagc cggcacgacg acgacgacat ccgaggctgt
     3061 gcggtgctgg acctggcctc cctgcagtgg gtggccatgc agtgcgacac acagctggac
     3121 tggatctgca agatccccag aggtacggac gtgcgggagc ccgacgacag ccctcaaggc
     3181 cgacgggaat ggctgcgctt ccaggaggcc gagtacaagt tctttgagca ccactccacg
     3241 tgggcgcagg cgcagcgcat ctgcacgtgg ttccaggccg agctgacctc ggtgcacagc
     3301 caggcggagc tagacttcct gagccacaac ttgcagaagt tctcccgggc ccaggagcag
     3361 cactggtgga tcggcctgca cacctctgag agcgatgggc gcttcagatg gacagatggt
     3421 tccattataa acttcatctc ctgggcacca ggcaaacctc ggcctgtcgg caaggacaag
     3481 aagtgcgtgt acatgacagc cagccgagag gactgggggg accagaggtg cctgacagcc
     3541 ttgccctaca tctgcaagcg cagcaacgtc accaaagaaa cgcagccccc agacctgcca
     3601 actacagccc tggggggctg cccctctgac tggatccagt tcctcaacaa gtgttttcag
     3661 gtccagggcc aggaacccca gagccgggtg aagtggtcag aggcacagtt ctcctgtgaa
     3721 cagcaagagg cccagctggt caccatcaca aaccccttag agcaagcatt catcacagcc
     3781 agcctgccca atgtgacctt tgacctttgg attggcctcc atgcctcgca gagggacttc
     3841 cagtgggtgg agcaggagcc tttgatgtat gccaactggg cacctgggga gccctctggc
     3901 cctagccctg ctcccagtgg caacaaaccg accagctgtg cagtggtcct gcacagcccc
     3961 tcagcccact tcactggccg ctgggacgat cggagctgca cggaggagac ccatggcttc
     4021 atctgccaga agggcacgga cccctccctg agcccgtccc cagcagcgct gccccccgcc
     4081 ccgggcactg agctctccta cctcaacggc accttccggc tgcttcagaa gccgctgcgc
     4141 tggcacgatg ccctcctgct gtgtgagagc cgcaatgcca gcctggccta cgtgcccgac
     4201 ccctacaccc aggccttcct cacgcaggct gcccgagggc tgcgcacgcc gctctggatt
     4261 gggctggctg gcgaggaggg ctctcggcgg tactcctggg tctcagagga gccgctgaac
     4321 tacgtgggct ggcaggacgg ggagccgcag cagccggggg gctgtaccta cgtagatgtg
     4381 gacggggcct ggcgcaccac cagctgtgac accaagctgc agggggctgt gtgtggggtt
     4441 agcagtgggc cccctcctcc ccgaagaata agctaccatg gcagctgtcc ccagggactg
     4501 gcagactccg cgtggattcc cttccgggag cactgctatt ctttccacat ggagctgctg
     4561 ctgggccaca aggaggcgcg acagcgctgc cagagagcgg gtggggccgt cctgtctatc
     4621 ctggatgaga tggagaatgt gtttgtctgg gagcacctgc agagctatga gggccagagt
     4681 cggggcgcct ggctgggcat gaacttcaac cccaaaggag gcactctggt ctggcaggac
     4741 aacacagctg tgaactactc caactggggg cccccgggct tgggccccag catgctgagc
     4801 cacaacagct gctactggat tcagagcaac agcgggctat ggcgccccgg cgcttgcacc
     4861 aacatcacca tgggtgtcgt ctgcaagctt cctcgtgctg agcagagcag cttctcccca
     4921 tcagcgcttc cagagaaccc agcggccctg gtggtggtgc tgatggcggt gctgctgctc
     4981 ctggccttgc tgaccgcagc cctcatcctt taccggaggc gccagagcat cgagcgcggg
     5041 gcctttgagg gtgcccgcta cagccgcagc agctccagcc ccaccgaggc cactgagaag
     5101 aacatcctgg tgtcagacat ggaaatgaat gagcaacaag aactcgagga acaaaaacta
     5161 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     5221 atttccgagg aggacttata gtgagggccc gatatctcga caatcaacct ctggattaca
     5281 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5341 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5401 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5461 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5521 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5581 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5641 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5701 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5761 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5821 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     5881 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5941 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6001 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6061 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6121 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     6181 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6241 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6301 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     6361 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6421 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6481 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     6541 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     6601 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6661 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6781 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6841 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6901 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6961 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     7021 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     7081 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     7141 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     7201 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     7261 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7321 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7381 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7441 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7501 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     7561 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7621 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7681 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7741 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7801 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     7861 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7921 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     7981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8101 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     8161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8221 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8281 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8521 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     8761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     9001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     9121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     9181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     9301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    10021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    10081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    10141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    10201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    10261 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
    10321 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
    10381 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
    10441 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
