LOCUS       V046429                 7087 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046429
VERSION     V046429
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2523
                     /label="PKM(NM_002654)"
                     /note="PKM(NM_002654)"
                     /gene="PKM"
     CDS             2530..2619
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2654..2878
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2924..3352
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3366..3695
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3762..4556
                     /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    4730..4851
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4900..4916)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4924..4940
                     /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(4948..4978)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4993..5014
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5302..5887)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6058..6918)
                     /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(6919..7023)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtcgaagc cccatagtga agccgggact
      961 gccttcattc agacccagca gctgcacgca gccatggctg acacattcct ggagcacatg
     1021 tgccgcctgg acattgattc accacccatc acagcccgga acactggcat catctgtacc
     1081 attggcccag cttcccgatc agtggagacg ttgaaggaga tgattaagtc tggaatgaat
     1141 gtggctcgtc tgaacttctc tcatggaact catgagtacc atgcggagac catcaagaat
     1201 gtgcgcacag ccacggaaag ctttgcttct gaccccatcc tctaccggcc cgttgctgtg
     1261 gctctagaca ctaaaggacc tgagatccga actgggctca tcaagggcag cggcactgca
     1321 gaggtggagc tgaagaaggg agccactctc aaaatcacgc tggataacgc ctacatggaa
     1381 aagtgtgacg agaacatcct gtggctggac tacaagaaca tctgcaaggt ggtggaagtg
     1441 ggcagcaaga tctacgtgga tgatgggctt atttctctcc aggtgaagca gaaaggtgcc
     1501 gacttcctgg tgacggaggt ggaaaatggt ggctccttgg gcagcaagaa gggtgtgaac
     1561 cttcctgggg ctgctgtgga cttgcctgct gtgtcggaga aggacatcca ggatctgaag
     1621 tttggggtcg agcaggatgt tgatatggtg tttgcgtcat tcatccgcaa ggcatctgat
     1681 gtccatgaag ttaggaaggt cctgggagag aagggaaaga acatcaagat tatcagcaaa
     1741 atcgagaatc atgagggggt tcggaggttt gatgaaatcc tggaggccag tgatgggatc
     1801 atggtggctc gtggtgatct aggcattgag attcctgcag agaaggtctt ccttgctcag
     1861 aagatgatga ttggacggtg caaccgagct gggaagcctg tcatctgtgc tactcagatg
     1921 ctggagagca tgatcaagaa gccccgcccc actcgggctg aaggcagtga tgtggccaat
     1981 gcagtcctgg atggagccga ctgcatcatg ctgtctggag aaacagccaa aggggactat
     2041 cctctggagg ctgtgcgcat gcagcacctg attgcccgtg aggcagaggc tgccatctac
     2101 cacttgcaat tatttgagga actccgccgc ctggcgccca ttaccagcga ccccacagaa
     2161 gccaccgccg tgggtgccgt ggaggcctcc ttcaagtgct gcagtggggc cataatcgtc
     2221 ctcaccaagt ctggcaggtc tgctcaccag gtggccagat accgcccacg tgcccccatc
     2281 attgctgtga cccggaatcc ccagacagct cgtcaggccc acctgtaccg tggcatcttc
     2341 cctgtgctgt gcaaggaccc agtccaggag gcctgggctg aggacgtgga cctccgggtg
     2401 aactttgcca tgaatgttgg caaggcccga ggcttcttca agaagggaga tgtggtcatt
     2461 gtgctgaccg gatggcgccc tggctccggc ttcaccaaca ccatgcgtgt tgttcctgtg
     2521 ccgctcgagg aacaaaaact aatatcagag gaagatttgg aacagaaact aatcagtgaa
     2581 gaagatttag agcaaaagtt aatttccgag gaggacttat gagggcccgt ttaaacccgc
     2641 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2701 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2761 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2821 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2881 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4741 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4801 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4861 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5821 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7021 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7081 ctatggt
//
