LOCUS       V039711                 7953 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039711
VERSION     V039711
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..2633
                     /label="CRAT(NM_000755)"
                     /note="CRAT(NM_000755)"
                     /gene="CRAT"
     CDS             2640..2705
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2729..3317
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3383..3431
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3633..4061
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4075..4404
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4471..5265
                     /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    5441..5562
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5611..5627)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5635..5651
                     /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(5659..5689)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5704..5725
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6772..7632)
                     /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(7633..7737)
                     /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 ccaccatgtt agccttcgct gccaggaccg
      781 tggtgaagcc tctgggcttc ctgaagccct tctccttgat gaaggcttcc agccgcttca
      841 aggcacacca ggatgcactg ccacggctgc ccgtgccccc tctccagcag tccctggacc
      901 actacctgaa ggcgctgcag cccatcgtga gtgaggagga gtgggcccac accaagcagc
      961 tggtggatga gtttcaggcc tcaggaggtg taggggagcg cctgcagaag gggctggagc
     1021 gtcgggccag gaagacggag aactggctgt ctgagtggtg gctcaagacc gcctacctcc
     1081 agtaccgcca gcctgtggtc atctactcga gcccaggcgt gatgctaccc aagcaggact
     1141 tcgtggacct gcagggtcag ctccgatttg ctgccaaact cattgagggt gtgttggatt
     1201 tcaaggtcat gattgacaac gagaccctgc ccgtggagta cctggggggg aagccactgt
     1261 gcatgaacca gtactatcag atcttgtcct cctgccgagt gccgggcccc aagcaggaca
     1321 cagtcagcaa cttcagcaag accaagaagc ctcccacgca catcaccgtg gtacacaact
     1381 accagttttt tgagctggat gtgtaccaca gtgacgggac acccctcact gcggatcaga
     1441 tctttgtgca gctggagaag atctggaact catccctaca gaccaacaag gagcctgtgg
     1501 gcatcctcac ctccaaccac cgcaactcct gggccaaggc atacaacacc ctcatcaaag
     1561 acaaggtgaa ccgggattcc gtgcgctcca tccagaagag catcttcacc gtgtgcctag
     1621 atgcaaccat gcccagggtc tcagaagacg tgtaccgcag ccacgtggca ggccagatgc
     1681 tgcatggggg cggcagcagg ctcaacagcg gcaaccgctg gttcgacaag acgctgcagt
     1741 tcatcgtggc agaagatggc tcctgtgggc ttgtgtacga gcatgctgca gcggaggggc
     1801 cccctattgt cacccttctg gactatgtca tcgagtacac gaagaaaccc gagcttgtgc
     1861 ggtctcccct ggtgcccctg cccatgccca agaagctgcg gttcaacatc acccccgaga
     1921 tcaagagcga catcgagaag gccaagcaga acctcagcat catgatccag gacctggata
     1981 tcaccgtgat ggtgttccac cattttggaa aagacttccc caagtcggag aagctaagcc
     2041 cagatgcctt catccagatg gctttgcagc tggcctacta caggatctac ggacaggcat
     2101 gtgccaccta tgaaagtgcc tccctgcgca tgtttcacct gggccgcacc gacaccatcc
     2161 gctcggcttc catggactca ctcacctttg tcaaggccat ggatgactcc agcgtcacgg
     2221 agcaccagaa ggtggagctg ctgcggaagg ccgtgcaggc ccaccgaggc tacaccgacc
     2281 gggccatccg cggggaggcc tttgatcgac acctgctggg cctgaagctg caggccatcg
     2341 aggacctggt gagcatgccc gacatcttca tggacacctc ctacgccatc gccatgcact
     2401 tccacctctc caccagccag gtccctgcca agacagactg tgtcatgttc ttcgggcccg
     2461 tggtccccga cggctacggt gtctgctata accccatgga ggcccacatc aacttctccc
     2521 tgtcggccta caacagctgc gcggagacca acgccgcccg cctggcgcat tacctggaga
     2581 aggcgctcct ggacatgcgt gccctgctgc agagccaccc ccgggccaag ctcggatccg
     2641 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     2701 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2761 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2821 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2881 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2941 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3001 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3061 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3121 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3181 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3241 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3301 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3361 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3421 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3481 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3541 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     3601 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3781 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3841 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3961 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4081 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4201 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4261 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4321 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4381 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4441 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4501 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4561 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4621 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4681 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4741 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4801 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4861 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4921 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4981 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5041 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5101 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5161 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5221 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5281 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5341 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5401 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5581 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5641 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5701 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5761 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6721 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6781 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6841 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6901 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6961 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7021 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7081 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7141 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7201 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7261 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7321 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7381 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7441 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7501 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7561 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7621 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7741 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7801 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7861 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7921 ttgaccgaca attgcatgaa gaatctgctt agg
//
