LOCUS       V026230                 7866 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026230
VERSION     V026230
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..2546
                     /label="F12(NM_021489)"
                     /note="F12(NM_021489)"
                     /gene="F12"
     CDS             2553..2618
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2642..3230
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3296..3344
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3546..3974
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3988..4317
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4384..5178
                     /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    5354..5475
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5524..5540)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5548..5564
                     /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(5572..5602)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5617..5638
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5926..6514)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6685..7545)
                     /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(7546..7650)
                     /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 ccaccatgac ggctctgttg ttcctggggt
      781 ctctgctgat gagtctggat ctgacacttt cggctccacc atggaaagac tccaagaaat
      841 ttaaggacgc acctgatggg cccacagtgg ttctcactgt ggatgggagg ctctgccatt
      901 ttccctttca gtaccaccgt cagctacacc acaaatgcat ccacaaaagg cggccaggct
      961 cccgcccctg gtgtgctacc acccccaact ttgatgaaga tcagcaatgg ggatactgct
     1021 tggagcccaa gaaagtgaaa gaccattgca gcaaacacaa cccgtgccac aaaggaggga
     1081 catgtatcaa cacccccaat gggccacact gtctctgccc tgaacacctc actgggaaac
     1141 attgccagaa agagaaatgc tttgagcctc agcttctcaa gttcttccac gagaatgagc
     1201 tatggtttag aacggggcca ggaggtgtgg ccaggtgcga gtgcaaaggt tctgaggctc
     1261 actgcaagcc ggtggccagc caggcctgca gcatcaatcc gtgccttaat gggggcagct
     1321 gcctcctcgt ggaggaccac ccactgtgcc gttgccctac aggctacact ggatattttt
     1381 gcgacttgga cctttgggcg acctgctatg aaggcagggg gctcagctac cggggccagg
     1441 ctggaactac gcaatcgggt gcgccatgtc agcggtggac cgtggaggcc acctaccgga
     1501 acatgactga gaagcaagcg ctaagctggg gcctgggcca ccacgcattt tgccggaacc
     1561 cagataatga cacacgtcca tggtgcttcg tctggagtgg cgacaggctg agctgggact
     1621 attgcggcct ggagcagtgc cagacgccaa cgtttgcacc tctagttgtc cctgagagtc
     1681 aggaggagtc cccgtcccag gcaccatctc tgtcccatgc accaaatgac tcgaccgatc
     1741 atcagacttc tctgtccaag accaacacga tgggctgcgg acagaggttc cgcaagggac
     1801 tgtcctcgtt catgcgcgtg gtgggcggac tagtggctct gcctgggtcg cacccctaca
     1861 tcgctgcact gtactggggt aacaacttct gcgcgggcag tctcatcgcc ccctgttggg
     1921 tgctgaccgc ggctcactgc ctgcagaatc ggccagcgcc cgaggaactg acagtggtac
     1981 ttggtcaaga tcgccacaac cagagctgcg agtggtgcca gactctggct gtgcgctcct
     2041 accgccttca cgagggcttc tcctccatca cctaccagca cgacttggct ctgctgcgcc
     2101 tgcaggaaag caaaaccaac agttgcgcga tcctgtcacc tcacgttcag cctgtgtgtc
     2161 tacccagcgg cgcggcccca ccctctgaga cagtgctctg cgaggtggcc ggctggggtc
     2221 accagttcga gggggctgaa gaatactcca ccttcctgca ggaggcacag gttcccttta
     2281 tcgccctgga tcgctgctcc aactctaacg tgcacggaga cgccattctc cctgggatgc
     2341 tttgcgctgg cttcttggag ggaggcaccg atgcctgcca gggtgactcc gggggccctc
     2401 tggtgtgtga ggaaggaact gcagaacatc agctcaccct gcgcggagtc atcagctggg
     2461 gctccggctg tggtgaccgc aacaagcccg gagtctacac agacgtggcc aactacctgg
     2521 cttggatcca gaagcatatt gcttcaggat ccgactacaa agaccatgac ggtgattata
     2581 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2641 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2701 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2761 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2821 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2881 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2941 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3001 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3061 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3121 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3181 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3241 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3301 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3361 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3421 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3481 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3541 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3901 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3961 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4081 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4141 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4201 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4261 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4321 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4381 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4441 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4501 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4561 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4621 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4681 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4741 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4801 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4861 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4921 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4981 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5041 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5101 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5161 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5221 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5281 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5341 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5401 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5461 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6121 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6361 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7681 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7741 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7801 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7861 cttagg
//
