LOCUS       V046507                 7386 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046507
VERSION     V046507
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..2066
                     /label="Krt13(NM_010662)"
                     /note="Krt13(NM_010662)"
                     /gene="Krt13"
     CDS             2073..2138
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2162..2750
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2816..2864
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3066..3494
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3508..3837
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3904..4698
                     /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    4874..4995
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5044..5060)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5068..5084
                     /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(5092..5122)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5137..5158
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5446..6034)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6205..7065)
                     /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(7066..7170)
                     /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 ccaccatgag ctgccgcttc cagagttcct
      781 ccatgagcta tggaggtggt ttcggagctg gttcttgcca gcttggagga ggccgtaata
      841 tctcctcctg ctcatctcgg tttgtcactg gagggtcagc tggaggctat ggagggggca
      901 tgagctgtgg ctttggtgga ggagctggtg gtggctttgg aggtggcttt ggaggtggct
      961 ttggaggtag ctatggcgga ggttttggcg gtggctttgg tgacttcgga ggtgtcgatg
     1021 gtggcctcct ctctggcaac gagaagatca ccatgcagaa cctcaatgac cgcctggcca
     1081 gctacctgga taaggtgcgc gccttggagg cagccaatgc cgacctggag gtgaagattc
     1141 gtgactggca tctgaaacag agcccagcta gtccagagcg ggactacagt gcttattata
     1201 agaccatcga ggagctccgg atcaagattc tggaagccac cacggacaac aaccggatca
     1261 tcctggagat tgacaatgcc aggctggctg cagatgactt caggctcaag tacgaaaatg
     1321 agctgacctt gcgccagagc gtggaggccg acatcaacgg cctgcgcagg gtgctggatg
     1381 agctgactct ggctaagact gacctggaaa tgcagattga gagcctgaat gaagaactgg
     1441 cttacctgaa gaagaaccac gaagaggaga tgaaggaatt cagcaaccag gtagtaggcc
     1501 aggtcaatgt ggagatggac gccacccctg gcattgatct gacccgtgtg ctggctgaga
     1561 tgagggagca gtatgaagcc ctggcagaga agaatcggag ggatgctgag gaatggttcc
     1621 agaccaagag tgcagagctg aacaaggaag tatcctccaa cgctgaaatg atccagacca
     1681 gcaagacaga gatcacagaa ctcaggcgca cactccaggg cctggagatt gagctgcagt
     1741 cccagctgag catgaaagcc ggcctggaaa gtacgttggc agagacagag tgccgctacg
     1801 ccctgcagct gcaacagatc caggggctca tcagcagcat cgaggctcag ctgagcgagc
     1861 tccgaagtga gatggagtgc cagaaccagg aatacaagat gctgttggac atcaaaacaa
     1921 ggctggaaca ggagatcgcc acctaccgca gcctgctaga gggccaggat gctaagatga
     1981 ccggcttcaa ctccggagga aataacacta ctacctccaa cgggtccccc tcctccaatt
     2041 ccggtcgccc agatttccga aagtatggat ccgactacaa agaccatgac ggtgattata
     2101 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2161 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2221 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2281 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2341 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2401 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2461 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2521 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2581 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2641 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2701 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     2761 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     2821 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     2881 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     2941 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3001 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3061 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3301 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3361 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3421 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3481 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3601 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3661 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3721 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3781 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3841 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3901 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3961 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4021 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4081 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4141 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4201 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4261 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4321 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4381 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4441 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4501 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4561 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4621 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4681 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4741 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4801 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4861 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4921 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4981 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5041 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5101 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5161 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5221 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5281 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5341 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5641 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5881 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6181 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6241 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6301 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6361 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6421 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6481 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6541 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6601 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6661 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6721 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6781 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6841 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6901 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6961 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7021 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7081 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7141 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7201 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7261 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7321 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7381 cttagg
//
