LOCUS       V046204                 8146 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046204
VERSION     V046204
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..2802
                     /label="GFPT1(NM_001244710)"
                     /note="GFPT1(NM_001244710)"
                     /gene="GFPT1"
     CDS             2809..2898
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2922..3510
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3576..3624
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3826..4254
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4268..4597
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4664..5458
                     /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    5634..5755
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5804..5820)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5828..5844
                     /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(5852..5882)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5897..5918
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6965..7825)
                     /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(7826..7930)
                     /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 ccaccatgtg tggtatattt
      721 gcttacttaa actaccatgt tcctcgaacg agacgagaaa tcctggagac cctaatcaaa
      781 ggccttcaga gactggagta cagaggatat gattctgctg gtgtgggatt tgatggaggc
      841 aatgataaag attgggaagc caatgcctgc aaaatccagc ttattaagaa gaaaggaaaa
      901 gttaaggcac tggatgaaga agttcacaag caacaagata tggatttgga tatagaattt
      961 gatgtacacc ttggaatagc tcatacccgt tgggcaacac atggagaacc cagtcctgtc
     1021 aatagccacc cccagcgctc tgataaaaat aatgaattta tcgttattca caatggaatc
     1081 atcaccaact acaaagactt gaaaaagttt ttggaaagca aaggctatga cttcgaatct
     1141 gaaacagaca cagagacaat tgccaagctc gttaagtata tgtatgacaa tcgggaaagt
     1201 caagatacca gctttactac cttggtggag agagttatcc aacaattgga aggtgctttt
     1261 gcacttgtgt ttaaaagtgt tcattttccc gggcaagcag ttggcacaag gcgaggtagc
     1321 cctctgttga ttggtgtacg gagtgaacat aaactttcta ctgatcacat tcctatactc
     1381 tacagaacag ctaggactca gattggatca aaattcacac ggtggggatc acagggagaa
     1441 agaggcaaag acaagaaagg aagctgcaat ctctctcgtg tggacagcac aacctgcctt
     1501 ttcccggtgg aagaaaaagc agtggagtat tactttgctt ctgatgcaag tgctgtcata
     1561 gaacacacca atcgcgtcat ctttctggaa gatgatgatg ttgcagcagt agtggatgga
     1621 cgtctttcta tccatcgaat taaacgaact gcaggagatc accccggacg agctgtgcaa
     1681 acactccaga tggaactcca gcagatcatg aagggcaact tcagttcatt tatgcagaag
     1741 gaaatatttg agcagccaga gtctgtcgtg aacacaatga gaggaagagt caactttgat
     1801 gactatactg tgaatttggg tggtttgaag gatcacataa aggagatcca gagatgccgg
     1861 cgtttgattc ttattgcttg tggaacaagt taccatgctg gtgtagcaac acgtcaagtt
     1921 cttgaggagc tgactgagtt gcctgtgatg gtggaactag caagtgactt cctggacaga
     1981 aacacaccag tctttcgaga tgatgtttgc tttttcctta gtcaatcagg tgagacagca
     2041 gatactttga tgggtcttcg ttactgtaag gagagaggag ctttaactgt ggggatcaca
     2101 aacacagttg gcagttccat atcacgggag acagattgtg gagttcatat taatgctggt
     2161 cctgagattg gtgtggccag tacaaaggct tataccagcc agtttgtatc ccttgtgatg
     2221 tttgccctta tgatgtgtga tgatcggatc tccatgcaag aaagacgcaa agagatcatg
     2281 cttggattga aacggctgcc tgatttgatt aaggaagtac tgagcatgga tgacgaaatt
     2341 cagaaactag caacagaact ttatcatcag aagtcagttc tgataatggg acgaggctat
     2401 cattatgcta cttgtcttga aggggcactg aaaatcaaag aaattactta tatgcactct
     2461 gaaggcatcc ttgctggtga attgaaacat ggccctctgg ctttggtgga taaattgatg
     2521 cctgtgatca tgatcatcat gagagatcac acttatgcca agtgtcagaa tgctcttcag
     2581 caagtggttg ctcggcaggg gcggcctgtg gtaatttgtg ataaggagga tactgagacc
     2641 attaagaaca caaaaagaac gatcaaggtg ccccactcag tggactgctt gcagggcatt
     2701 ctcagcgtga tccctttaca gttgctggct ttccaccttg ctgtgctgag aggctatgat
     2761 gttgatttcc cacggaatct tgccaaatct gtgactgtag agctcgagga acaaaaacta
     2821 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     2881 atttccgagg aggacttata gtgagggccc gatatctcga caatcaacct ctggattaca
     2941 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3001 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3061 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3121 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3181 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3241 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3301 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3361 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3421 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3481 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     3541 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3601 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3661 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3721 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     3781 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3841 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3901 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3961 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     4021 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4081 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4141 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     4201 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4261 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4321 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4441 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4501 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4561 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4621 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4681 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4741 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4801 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4861 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4921 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4981 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5041 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5101 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5161 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5221 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5281 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5341 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5401 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5461 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5521 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5581 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5641 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5701 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5761 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7981 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8041 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8101 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
