LOCUS       V043769                 7596 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043769
VERSION     V043769
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..2276
                     /label="SLC2A6(NM_017585)"
                     /note="SLC2A6(NM_017585)"
                     /gene="SLC2A6"
     CDS             2283..2348
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2372..2960
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3026..3074
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3276..3704
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3718..4047
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4114..4908
                     /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    5084..5205
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5254..5270)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5278..5294
                     /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(5302..5332)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5347..5368
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5656..6244)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6415..7275)
                     /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(7276..7380)
                     /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 ccaccatgca ggagccgctg ctgggagccg
      781 agggcccgga ctacgacacc ttccccgaga agccgccccc gtcgccaggg gacagggcgc
      841 gggtcgggac cctgcagaac aaaagggtgt tcctggccac cttcgccgca gtgctcggca
      901 atttcagctt tgggtatgcc ctggtctaca catcccctgt catcccagcc ctggagcgct
      961 ccttggatcc tgacctgcat ctgaccaaat cccaggcatc ctggtttggg tccgtgttca
     1021 ccctgggagc agcggccgga ggcctgagtg ccatgatcct caacgacctc ctgggccgga
     1081 agctgagcat catgttctca gctgtgccgt cggcggccgg ctatgcgctc atggcgggtg
     1141 cgcacggcct ctggatgctg ctgctcggaa ggacgctgac gggcttcgcc ggggggctca
     1201 cagctgcctg catcccggtg tacgtgtctg agattgctcc cccaggcgtt cgtggggctc
     1261 tgggggccac accccagctc atggcagtgt tcggatccct gtccctctac gcccttggcc
     1321 tcctgctgcc gtggcgctgg ctggctgtgg ccggggaggc gcctgtgctc atcatgatcc
     1381 tgctgctcag cttcatgccc aactcgccgc gcttcctgct ctctcggggc agggacgaag
     1441 aggccctgcg ggcgctggcc tggctgcgtg ggacggacgt cgatgtccac tgggagttcg
     1501 agcagatcca ggacaacgtc cggagacaga gcagccgagt atcgtgggct gaggcacggg
     1561 ccccacacgt gtgccggccc atcaccgtgg ccttgctgat gcgcctcctg cagcagctga
     1621 cgggcatcac gcccatcctg gtctacctgc agtccatctt cgacagcacc gctgtcctgc
     1681 tgccccccaa ggacgacgca gccatcgttg gggccgtgcg gctcctgtcc gtgctgatcg
     1741 ccgccctcac catggacctc gcaggccgca aggtgctgct cttcgtctca gcggccatca
     1801 tgtttgctgc caacctgact ctggggctgt acatccactt tggccccagg cctctgagcc
     1861 ccaacagcac tgcgggcctg gaaagcgagt cctgggggga cttggcgcag cccctggcag
     1921 cacccgctgg ctacctcacc ctggtgcccc tgctggccac catgctcttc atcatgggct
     1981 acgccgtggg ctggggtccc atcacctggc tgctcatgtc tgaggtcctg cccctgcgtg
     2041 cccgtggcgt ggcctcaggg ctctgcgtgc tggccagctg gctcaccgcc ttcgtcctca
     2101 ccaagtcctt cctgccagtg gtgagcacct tcggcctcca ggtgcctttc ttcttcttcg
     2161 cggccatctg cttggtgagc ctggtgttca caggctgctg tgtgcccgag accaagggac
     2221 ggtccctgga gcagatcgag tccttcttcc gcacggggag aaggtccttc ttgcgcggat
     2281 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     2341 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     2401 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2461 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2521 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2581 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2641 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2701 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2761 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2821 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2881 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2941 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3001 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3061 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3121 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3181 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3241 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3421 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3601 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4921 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4981 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5041 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5101 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5161 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5221 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5281 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5341 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5401 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5461 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5521 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5581 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5641 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5701 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5761 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5821 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5881 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5941 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6001 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6061 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6121 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6181 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6241 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6301 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6361 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6421 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6481 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6541 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6601 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6661 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6721 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6781 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6841 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6901 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6961 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7021 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7081 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7141 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7201 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7261 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7321 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7381 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7441 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7501 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7561 ggcttgaccg acaattgcat gaagaatctg cttagg
//
