LOCUS       V044702                 6994 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044702
VERSION     V044702
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..1650
                     /label="Ddrgk1(NM_029832)"
                     /note="Ddrgk1(NM_029832)"
                     /gene="Ddrgk1"
     CDS             1657..1746
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1770..2358
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2424..2472
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2674..3102
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3116..3445
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3512..4306
                     /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    4482..4603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4652..4668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4676..4692
                     /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(4700..4730)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4745..4766
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5813..6673)
                     /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(6674..6778)
                     /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 ccaccatggt ggggccctgg
      721 gtgtatctgg tggcggcagt tttgctcatc ggcctgatcc tcttcctgac tcgcagccgg
      781 ggtcgggcgg cagcagctga cggagaacca ctgcacaatg aggaagagag ggcaggagca
      841 ggccaggtag gccgctcttt gccccaggag tctgaagaac agagaactgg aagcagaccc
      901 cggcgtcgga gggacttggg cagccgtcta caggcccagc gtcgagccca gcgagtggcc
      961 tgggaagacg gggatgagaa tgtgggtcaa actgttattc cagcccagga ggaagaaggc
     1021 attgagaagc cagcagaagt tcacccaaca gggaaaattg gagccaagaa actacggaag
     1081 ctagaggaaa aacaggctcg aaaggctcag cgagaggcag aggaggctga acgtgaagaa
     1141 cggaaacgcc tagagtccca acgtgaggcc gaatggaaga aggaagagga acggcttcgc
     1201 ctgaaggaag aacagaagga ggaggaagag aggaaggctc aggaggagca ggcccggcgg
     1261 gagcacgagg agtacctgaa actgaaggag gccttcgtgg tagaagaaga aggtgttagc
     1321 gaaaccatga ctgaggagca gtctcacagc ttcctgacag aattcatcaa ttacatcaag
     1381 aagtccaagg ttgtgctttt ggaagatctg gctttccaga tgggcctaag gactcaggac
     1441 gccataaacc gcatccagga cctgctgacg gaggggactc taacaggtgt gattgacgac
     1501 cggggcaagt ttatctacat aaccccagag gaactggctg ccgtggccaa tttcatccga
     1561 cagcggggcc gggtgtccat cacagagctt gcccaggcca gcaactccct catctcctgg
     1621 ggccaggacc tccctgccca ggcttcagcc ctcgaggaac aaaaactaat atcagaggaa
     1681 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     1741 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     1801 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1861 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1921 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     1981 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2041 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2101 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2161 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2221 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2281 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2341 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     2401 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     2461 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2521 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2581 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     2641 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2821 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2881 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3001 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     3061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3121 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3481 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3541 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3601 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3661 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3721 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3781 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3841 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3901 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3961 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4021 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4081 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4141 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4201 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4261 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4321 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4501 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4561 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6781 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6841 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6901 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6961 cttgaccgac aattgcatga agaatctgct tagg
//
