LOCUS       V027606                 8800 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027606
VERSION     V027606
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..3456
                     /label="HK2(NM_000189)"
                     /note="HK2(NM_000189)"
                     /gene="HK2"
     CDS             3463..3552
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3576..4164
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4230..4278
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4480..4908
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4922..5251
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5318..6112
                     /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    6288..6409
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6458..6474)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6482..6498
                     /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(6506..6536)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6551..6572
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6860..7448)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7619..8479)
                     /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(8480..8584)
                     /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 ccaccatgat tgcctcgcat
      721 ctgcttgcct acttcttcac ggagctcaac catgaccaag tgcagaaggt tgaccagtat
      781 ctctaccaca tgcgcctctc tgatgagacc ctcttggaga tctctaagcg gttccgcaag
      841 gagatggaga aagggcttgg agccaccact caccctactg cagcagtgaa gatgctgccc
      901 acctttgtga ggtccactcc agatgggaca gaacacggag agttcctggc tctggatctt
      961 ggagggacca acttccgtgt gctttgggtg aaagtaacgg acaatgggct ccagaaggtg
     1021 gagatggaga atcagatcta tgccatccct gaggacatca tgcgaggcag tggcacccag
     1081 ctgtttgacc acattgccga atgcctggct aacttcatgg ataagctaca aatcaaagac
     1141 aagaagctcc cactgggttt taccttctcg ttcccctgcc accagactaa actagacgag
     1201 agtttcctgg tctcatggac caagggattc aagtccagtg gagtggaagg cagagacgtt
     1261 gtggctctga tccggaaggc catccagagg agaggggact ttgatatcga cattgtggct
     1321 gtggtgaatg acacagttgg gaccatgatg acctgtggtt atgatgacca caactgtgag
     1381 attggtctca ttgtgggcac gggcagcaac gcctgctaca tggaagagat gcgccacatc
     1441 gacatggtgg aaggcgatga ggggcggatg tgtatcaata tggagtgggg ggccttcggg
     1501 gacgatggct cgctcaacga cattcgcact gagtttgacc aggagattga catgggctca
     1561 ctgaacccgg gaaagcaact gtttgagaag atgatcagtg ggatgtacat gggggagctg
     1621 gtgaggctta tcctggtgaa gatggccaag gaggagctgc tctttggggg gaagctcagc
     1681 ccagagcttc tcaacaccgg tcgctttgag accaaagaca tctcagacat tgaaggggag
     1741 aaggatggca tccggaaggc ccgtgaggtc ctgatgcggt tgggcctgga cccgactcag
     1801 gaggactgcg tggccactca ccggatctgc cagatcgtgt ccacacgctc cgccagcctg
     1861 tgcgcagcca ccctggccgc cgtgctgcag cgcatcaagg agaacaaagg cgaggagcgg
     1921 ctgcgctcta ctattggggt cgacggttcc gtctacaaga aacaccccca ttttgccaag
     1981 cgtctacata agaccgtgcg gcggctggtg cccggctgcg atgtccgctt cctccgctcc
     2041 gaggatggca gtggcaaagg tgcagccatg gtgacagcag tggcttaccg gctggccgat
     2101 caacaccgtg cccgccagaa gacattagag catctgcagc tgagccatga ccagctgctg
     2161 gaggtcaaga ggaggatgaa ggtagaaatg gagcgaggtc tgagcaagga gactcatgcc
     2221 agtgcccccg tcaagatgct gcccacctac gtgtgtgcta ccccggacgg cacagagaaa
     2281 ggggacttct tggccttgga ccttggagga acaaatttcc gggtcctgct ggtccgtgtt
     2341 cggaatggga agtggggtgg agtggagatg cacaacaaga tctacgccat cccgcaggag
     2401 gtcatgcacg gcaccgggga cgagctcttt gaccacattg tccagtgcat cgcggacttc
     2461 ctcgagtaca tgggcatgaa gggcgtgtcc ctgcctctgg gttttacctt ctccttcccc
     2521 tgccagcaga acagcctgga cgagagcatc ctcctcaagt ggacaaaagg cttcaaggca
     2581 tctggctgcg agggcgagga cgtggtgacc ctgctgaagg aagcgatcca ccggcgagag
     2641 gagtttgacc tggatgtggt tgctgtggtg aacgacacag tcggaactat gatgacctgt
     2701 ggctttgaag accctcactg tgaagttggc ctcattgttg gcacgggcag caatgcctgc
     2761 tacatggagg agatgcgcaa cgtggaactg gtggaaggag aagaggggcg gatgtgtgtg
     2821 aacatggaat ggggggcctt cggggacaat ggatgcctag atgacttccg cacagaattt
     2881 gatgtggctg tggatgagct ttcactcaac cccggcaagc agaggttcga gaaaatgatc
     2941 agtggaatgt acctgggtga gattgtccgt aacattctca tcgatttcac caagcgtgga
     3001 ctactcttcc gaggccgcat ctcagagcgg ctcaagacaa ggggcatctt tgaaaccaag
     3061 ttcttgtctc agattgagag tgactgcctg gccctgctgc aagtccgagc catcctgcaa
     3121 cacttagggc ttgagagcac ctgtgacgac agcatcattg ttaaggaggt gtgcactgtg
     3181 gtggcccggc gggcagccca gctctgtggc gcaggcatgg ccgctgtggt ggacaggata
     3241 cgagaaaacc gtgggctgga cgctctcaaa gtgacagtgg gtgtggatgg gaccctctac
     3301 aagctacatc ctcactttgc caaagtcatg catgagacag tgaaggacct ggctccgaaa
     3361 tgtgatgtgt ctttcctgca gtcagaggat ggcagcggga agggggcggc gctcatcact
     3421 gctgtggcct gccgcatccg tgaggctgga cagcgactcg aggaacaaaa actaatatca
     3481 gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa gttaatttcc
     3541 gaggaggact tatagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt
     3601 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3661 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3721 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3781 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3841 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3901 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3961 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4021 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4081 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4141 tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga
     4201 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4261 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4321 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4381 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4441 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4501 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4561 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4621 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4681 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4741 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4801 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     4861 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4921 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4981 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5041 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5101 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5161 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5221 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5281 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5341 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5401 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5461 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5521 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5581 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5641 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5701 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5761 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5821 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5881 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5941 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6001 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6061 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6121 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6181 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6241 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     6301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6421 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6481 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6541 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6601 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6661 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7021 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     7081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7261 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8581 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     8641 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     8701 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     8761 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
