LOCUS       V020256                 8159 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020256
VERSION     V020256
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3691
                     /label="HK1(NM_000188)"
                     /note="HK1(NM_000188)"
                     /gene="HK1"
     CDS             3698..3721
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    3759..3983
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4029..4457
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4471..4800
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4867..5661
                     /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    5835..5956
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6005..6021)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6029..6045
                     /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(6053..6083)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6098..6119
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6407..6992)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7163..8023)
                     /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(8024..8128)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgatcgccg cgcagctcct
      961 ggcctattac ttcacggagc tgaaggatga ccaggtcaaa aagattgaca agtatctcta
     1021 tgccatgcgg ctctccgatg aaactctcat agatatcatg actcgcttca ggaaggagat
     1081 gaagaatggc ctctcccggg attttaatcc aacagccaca gtcaagatgt tgccaacatt
     1141 cgtaaggtcc attcctgatg gctctgaaaa gggagatttc attgccctgg atcttggtgg
     1201 gtcttccttt cgaattctgc gggtgcaagt gaatcatgag aaaaaccaga atgttcacat
     1261 ggagtccgag gtttatgaca ccccagagaa catcgtgcac ggcagtggaa gccagctttt
     1321 tgatcatgtt gctgagtgcc tgggagattt catggagaaa aggaagatca aggacaagaa
     1381 gttacctgtg ggattcacgt tttcttttcc ttgccaacaa tccaaaatag atgaggccat
     1441 cctgatcacc tggacaaagc gatttaaagc gagcggagtg gaaggagcag atgtggtcaa
     1501 actgcttaac aaagccatca aaaagcgagg ggactatgat gccaacatcg tagctgtggt
     1561 gaatgacaca gtgggcacca tgatgacctg tggctatgac gaccagcact gtgaagtcgg
     1621 cctgatcatc ggcactggca ccaatgcttg ctacatggag gaactgaggc acattgatct
     1681 ggtggaagga gacgagggga ggatgtgtat caatacagaa tggggagcct ttggagacga
     1741 tggatcatta gaagacatcc ggacagagtt tgacagggag atagaccggg gatccctcaa
     1801 ccctggaaaa cagctgtttg agaagatggt cagtggcatg tacttgggag agctggttcg
     1861 actgatccta gtcaagatgg ccaaggaggg cctcttattt gaagggcgga tcaccccgga
     1921 gctgctcacc cgagggaagt ttaacaccag tgatgtgtca gccatcgaaa agaataagga
     1981 aggcctccac aatgccaaag aaatcctgac ccgcctggga gtggagccgt ccgatgatga
     2041 ctgtgtctca gtccagcacg tttgcaccat tgtctcattt cgctcagcca acttggtggc
     2101 tgccacactg ggcgccatct tgaaccgcct gcgtgataac aagggcacac ccaggctgcg
     2161 gaccacggtt ggtgtcgacg gatctcttta caagacgcac ccacagtatt cccggcgttt
     2221 ccacaagact ctaaggcgct tggtgccaga ctccgatgtg cgcttcctcc tctcggagag
     2281 tggcagcggc aagggggctg ccatggtgac ggcggtggcc taccgcttgg ccgagcagca
     2341 ccggcagata gaggagaccc tggctcattt ccacctcacc aaggacatgc tgctggaggt
     2401 gaagaagagg atgcgggccg agatggagct ggggctgagg aagcagacgc acaacaatgc
     2461 cgtggttaag atgctgccct ccttcgtccg gagaactccc gacgggaccg agaatggtga
     2521 cttcttggcc ctggatcttg gaggaaccaa tttccgtgtg ctgctggtga aaatccgtag
     2581 tgggaaaaag agaacggtgg aaatgcacaa caagatctac gccattccta ttgaaatcat
     2641 gcagggcact ggggaagagc tgtttgatca cattgtctcc tgcatctctg acttcttgga
     2701 ctacatgggg atcaaaggcc ccaggatgcc tctgggcttc acgttctcat ttccctgcca
     2761 gcagacgagt ctggacgcgg gaatcttgat cacgtggaca aagggtttta aggcaacaga
     2821 ctgcgtgggc cacgatgtag tcaccttact aagggatgcg ataaaaagga gagaggaatt
     2881 tgacctggac gtggtggctg tggtcaacga cacagtgggc accatgatga cctgtgctta
     2941 tgaggagccc acctgtgagg ttggactcat tgttgggacc ggcagcaatg cctgctacat
     3001 ggaggagatg aagaacgtgg agatggtgga gggggaccag gggcagatgt gcatcaacat
     3061 ggagtggggg gcctttgggg acaacgggtg tctggatgat atcaggacac actacgacag
     3121 actggtggac gaatattccc taaatgctgg gaaacaaagg tatgagaaga tgatcagtgg
     3181 tatgtacctg ggtgaaatcg tccgcaacat cttaatcgac ttcaccaaga agggattcct
     3241 cttccgaggg cagatctctg agacgctgaa gacccggggc atctttgaga ccaagtttct
     3301 ctctcagatc gagagtgacc gattagcact gctccaggtc cgggctatcc tccagcagct
     3361 aggtctgaat agcacctgcg atgacagtat cctcgtcaag acagtgtgcg gggtggtgtc
     3421 caggagggcc gcacagctgt gtggcgcagg catggctgcg gttgtggata agatccgcga
     3481 gaacagagga ctggaccgtc tgaatgtgac tgtgggagtg gacgggacac tctacaagct
     3541 tcatccacac ttctccagaa tcatgcacca gacggtgaag gaactgtcac caaaatgtaa
     3601 cgtgtccttc ctcctgtctg aggatggcag cggcaagggg gccgccctca tcacggccgt
     3661 gggcgtgcgg ttacgcacag aggcaagcag cctcgaggat tacaaggatg acgacgataa
     3721 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3781 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3841 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3901 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3961 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4021 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4081 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4141 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4201 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4261 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4321 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4381 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4441 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4501 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4681 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4741 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4801 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4861 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4921 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4981 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5041 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5101 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5161 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5221 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5281 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5341 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5401 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5461 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5521 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5581 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5641 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5701 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5761 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5821 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5881 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5941 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6001 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6061 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6121 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6181 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6241 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6301 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6361 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6421 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6481 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6541 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6601 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6661 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6721 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6781 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6841 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6901 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7081 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7141 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7201 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7261 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7321 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7381 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7441 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7501 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7561 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7621 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7681 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7741 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7801 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7861 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7921 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7981 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8041 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
