LOCUS       V026167                 8233 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026167
VERSION     V026167
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..3753
                     /label="HK1(NM_033497)"
                     /note="HK1(NM_033497)"
                     /gene="HK1"
     polyA_signal    3833..4057
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4103..4531
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4545..4874
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4941..5735
                     /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    5909..6030
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6079..6095)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6103..6119
                     /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(6127..6157)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6172..6193
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6481..7066)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7237..8097)
                     /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(8098..8202)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggggcaga tctgccagcg agaatcggct
     1021 acagcagctg aaaaaccaaa acttcatcta cttgctgaaa gtgagattga caagtatctc
     1081 tatgccatgc ggctctccga tgaaactctc atagatatca tgactcgctt caggaaggag
     1141 atgaagaatg gcctctcccg ggattttaat ccaacagcca cagtcaagat gttgccaaca
     1201 ttcgtaaggt ccattcctga tggctctgaa aagggagatt tcattgccct ggatcttggt
     1261 gggtcttcct ttcgaattct gcgggtgcaa gtgaatcatg agaaaaacca gaatgttcac
     1321 atggagtccg aggtttatga caccccagag aacatcgtgc acggcagtgg aagccagctt
     1381 tttgatcatg ttgctgagtg cctgggagat ttcatggaga aaaggaagat caaggacaag
     1441 aagttacctg tgggattcac gttttctttt ccttgccaac aatccaaaat agatgaggcc
     1501 atcctgatca cctggacaaa gcgatttaaa gcgagcggag tggaaggagc agatgtggtc
     1561 aaactgctta acaaagccat caaaaagcga ggggactatg atgccaacat cgtagctgtg
     1621 gtgaatgaca cagtgggcac catgatgacc tgtggctatg acgaccagca ctgtgaagtc
     1681 ggcctgatca tcggcactgg caccaatgct tgctacatgg aggaactgag gcacattgat
     1741 ctggtggaag gagacgaggg gaggatgtgt atcaatacag aatggggagc ctttggagac
     1801 gatggatcat tagaagacat ccggacagag tttgacaggg agatagaccg gggatccctc
     1861 aaccctggaa aacagctgtt tgagaagatg gtcagtggca tgtacttggg agagctggtt
     1921 cgactgatcc tagtcaagat ggccaaggag ggcctcttat ttgaagggcg gatcaccccg
     1981 gagctgctca cccgagggaa gtttaacacc agtgatgtgt cagccatcga aaagaataag
     2041 gaaggcctcc acaatgccaa agaaatcctg acccgcctgg gagtggagcc gtccgatgat
     2101 gactgtgtct cagtccagca cgtttgcacc attgtctcat ttcgctcagc caacttggtg
     2161 gctgccacac tgggcgccat cttgaaccgc ctgcgtgata acaagggcac acccaggctg
     2221 cggaccacgg ttggtgtcga cggatctctt tacaagacgc acccacagta ttcccggcgt
     2281 ttccacaaga ctctaaggcg cttggtgcca gactccgatg tgcgcttcct cctctcggag
     2341 agtggcagcg gcaagggggc tgccatggtg acggcggtgg cctaccgctt ggccgagcag
     2401 caccggcaga tagaggagac cctggctcat ttccacctca ccaaggacat gctgctggag
     2461 gtgaagaaga ggatgcgggc cgagatggag ctggggctga ggaagcagac gcacaacaat
     2521 gccgtggtta agatgctgcc ctccttcgtc cggagaactc ccgacgggac cgagaatggt
     2581 gacttcttgg ccctggatct tggaggaacc aatttccgtg tgctgctggt gaaaatccgt
     2641 agtgggaaaa agagaacggt ggaaatgcac aacaagatct acgccattcc tattgaaatc
     2701 atgcagggca ctggggaaga gctgtttgat cacattgtct cctgcatctc tgacttcttg
     2761 gactacatgg ggatcaaagg ccccaggatg cctctgggct tcacgttctc atttccctgc
     2821 cagcagacga gtctggacgc gggaatcttg atcacgtgga caaagggttt taaggcaaca
     2881 gactgcgtgg gccacgatgt agtcacctta ctaagggatg cgataaaaag gagagaggaa
     2941 tttgacctgg acgtggtggc tgtggtcaac gacacagtgg gcaccatgat gacctgtgct
     3001 tatgaggagc ccacctgtga ggttggactc attgttggga ccggcagcaa tgcctgctac
     3061 atggaggaga tgaagaacgt ggagatggtg gagggggacc aggggcagat gtgcatcaac
     3121 atggagtggg gggcctttgg ggacaacggg tgtctggatg atatcaggac acactacgac
     3181 agactggtgg acgaatattc cctaaatgct gggaaacaaa ggtatgagaa gatgatcagt
     3241 ggtatgtacc tgggtgaaat cgtccgcaac atcttaatcg acttcaccaa gaagggattc
     3301 ctcttccgag ggcagatctc tgagacgctg aagacccggg gcatctttga gaccaagttt
     3361 ctctctcaga tcgagagtga ccgattagca ctgctccagg tccgggctat cctccagcag
     3421 ctaggtctga atagcacctg cgatgacagt atcctcgtca agacagtgtg cggggtggtg
     3481 tccaggaggg ccgcacagct gtgtggcgca ggcatggctg cggttgtgga taagatccgc
     3541 gagaacagag gactggaccg tctgaatgtg actgtgggag tggacgggac actctacaag
     3601 cttcatccac acttctccag aatcatgcac cagacggtga aggaactgtc accaaaatgt
     3661 aacgtgtcct tcctcctgtc tgaggatggc agcggcaagg gggccgccct catcacggcc
     3721 gtgggcgtgc ggttacgcac agaggcaagc agctaagaat tctgcagata tccagcacag
     3781 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3841 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3901 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3961 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4021 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4081 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4141 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4201 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4261 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4321 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4381 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4441 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4501 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4561 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4801 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4861 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4921 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4981 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5041 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5101 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5161 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5221 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5281 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5341 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5401 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5461 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5521 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5581 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5641 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5701 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5761 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5821 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5881 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5941 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6001 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6061 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     7021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8101 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8161 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8221 gccacctgac gtc
//
