LOCUS       V040315                 9646 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040315
VERSION     V040315
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..4302
                     /label="TTLL4(NM_014640)"
                     /note="TTLL4(NM_014640)"
                     /gene="TTLL4"
     CDS             4309..4398
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4422..5010
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5076..5124
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5326..5754
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5768..6097
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6164..6958
                     /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    7134..7255
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7304..7320)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7328..7344
                     /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(7352..7382)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7397..7418
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7706..8294)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8465..9325)
                     /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(9326..9430)
                     /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 ccaccatggc ctcagcagga
      721 acacagcact atagtattgg cctccgccag aaaaacagct tcaagcagag tggtccctca
      781 ggcacagtac ctgccacgcc acctgagaaa ccctcggagg gcagagtctg gcctcaggcc
      841 catcagcaag tgaagccaat ctggaagctg gaaaagaagc aagtggagac actgtcagca
      901 gggttgggcc caggcctctt gggcgtccca ccccagccag catatttctt ttgccccagc
      961 actttatgta gctctgggac cacggctgtc attgcaggcc acagcagttc ctgttaccta
     1021 cactctctcc cggacttgtt caacagcacc ctgctatacc gccgctccag ctataggcaa
     1081 aaaccgtacc agcaactgga gtctttctgc ttgcgttcga gcccgtcaga aaaaagccct
     1141 ttttctctcc ctcaaaagag cctccctgtc agtctcactg ccaacaaggc cacttcttcc
     1201 atggtcttct ccatggccca gcccatggcc tcctcatcca cagaaccata cctctgcttg
     1261 gcagcggctg gggaaaaccc ttcagggaag agcctggcct ctgccatctc agggaagatc
     1321 ccatctccac tctcttcctc ctataagccc atgctgaata ataattcctt catgtggcca
     1381 aatagcacgc cagtgccttt attgcagacc acacagggcc tgaagccagt atcgccaccc
     1441 aagatccagc ctgtctcctg gcatcattca gggggtactg gagactgtgc accgcagcct
     1501 gttgaccata aggtgcccaa aagcattggc actgtcccag ctgatgccag tgcccatatc
     1561 gccttgtcta ccgctagctc ccacgacaca tccaccacca gtgttgcctc ttcctggtat
     1621 aaccggaata acttagccat gagggcagag ccactttcct gtgctctgga tgacagctct
     1681 gattcccagg atccaactaa ggagattcgg ttcactgagg ccgtgaggaa attgaccgca
     1741 agaggctttg agaagatgcc gaggcaaggc tgccagcttg aacagtctag tttcctgaac
     1801 cccagcttcc agtggaatgt cctcaacagg agcaggcggt ggaaacctcc tgcggtaaat
     1861 cagcagtttc ctcaggagga tgctggatcg gtcaggcggg tcctccctgg tgcctcagat
     1921 accttggggt tggacaatac agtcttctgt accaagcgta tcagcattca cctccttgcc
     1981 tcacatgcca gtgggctcaa tcacaaccct gcctgtgaat ctgtaattga ctcctcagca
     2041 tttggagaag gcaaagctcc aggtccccct tttcctcaaa ctcttggcat agccaacgtg
     2101 gccacccgcc tctcttccat ccagctgggc cagtctgaga aggagagacc tgaggaggcc
     2161 agggagctgg actcatctga tagggatatt agttcagcta ctgacctcca gccagatcag
     2221 gctgagactg aagatacaga agaagaacta gtagatggtt tggaagactg ttgtagccgt
     2281 gatgagaatg aagaggagga gggagactca gagtgctcct cattaagtgc tgtctccccc
     2341 agcgaatcgg tggccatgat ctctagaagc tgtatggaaa ttctgaccaa acccctttcc
     2401 aatcatgaga aagttgtccg accagccctc atctacagtc tctttcccaa cgttccccct
     2461 accatctatt ttggcactcg ggatgagaga gtggagaaac ttccctggga acagaggaag
     2521 ttgctccgat ggaagatgag cacagtgacc cccaacattg tcaagcagac cattggacgg
     2581 tcccacttca aaatcagcaa aagaaacgat gactggctgg gctgctgggg tcaccacatg
     2641 aagtctccta gtttccgatc cattcgagag catcagaagc taaaccattt cccaggctca
     2701 ttccagattg ggaggaagga ccggctatgg cggaacctgt cacgtatgca gagccgcttt
     2761 ggcaagaagg agttcagttt cttcccccag tcctttatcc tgccccagga cgccaagctc
     2821 ctgcgcaaag cgtgggagag cagcagccgc caaaagtgga ttgtgaagcc accagcatca
     2881 gctcgaggca ttggcatcca ggttattcac aagtggagtc agctccccaa gcgaaggccc
     2941 ctcctggtac agaggtatct acacaaaccc tacctcatca gcggcagcaa gtttgacctg
     3001 cggatctatg tttatgtcac ttcctacgat cctctgcgga tttacctctt ttcagatgga
     3061 ctggtccgct ttgccagttg caagtattcg ccttccatga agagccttgg caataagttc
     3121 atgcacctga ccaactacag tgtcaataaa aagaatgccg agtaccaggc caatgcagat
     3181 gaaatggctt gccagggcca caaatgggca ctgaaggctt tgtggaacta cctgagccag
     3241 aagggagtca atagcgacgc catctgggag aagataaagg atgttgttgt caaaactatc
     3301 atctcgtcag agccctatgt gaccagcctg ctcaagatgt atgtgcgacg gccctatagc
     3361 tgccatgaac tctttggttt tgacatcatg ctagacgaaa acctcaagcc ctgggtcctg
     3421 gaagtcaaca tttccccaag cctccactcc agctctccac tggatatcag catcaaaggc
     3481 cagatgattc gtgaccttct gaatctggca ggttttgtcc tgcccaatgc agaggatatc
     3541 atttccagcc ccagcagctg cagcagctcc accaccagcc tgcccacctc ccctggggac
     3601 aaatgtcgaa tggctccaga gcatgtcact gcacagaaga tgaagaaagc ctattatctg
     3661 acccagaaaa ttcctgatca ggacttctat gcatctgtgc tggatgtcct gacaccagat
     3721 gatgttcgga ttctggttga gatggaagat gagttttctc gccgtggtca gtttgaacga
     3781 atttttcctt ctcatatctc ctctcgctat ctccgctttt ttgagcagcc acgatatttc
     3841 aacattctca ccacccaatg ggaacagaaa taccatggca acaagcttaa aggagtagat
     3901 ctgctccgga gttggtgcta caaagggttc cacatgggag ttgtctctga ttctgctcca
     3961 gtgtggtctc tcccgacatc acttctgact atctcaaagg atgacgtgat actcaatgcc
     4021 ttcagcaaat cagagactag caagctggga aaacaaagct cctgtgaggt tagcctacta
     4081 ctctctgaag acgggaccac gcccaaatcc aagaagactc aagctggcct ttccccttat
     4141 ccccagaaac ccagttcctc aaaggacagt gaggacacca gcaaagagcc cagcctttct
     4201 acccagacgt tacctgtgat caagtgctct gggcagactt caagactttc tgcttcctcc
     4261 actttccagt caatcagtga ctccctcctg gctgtgagcc cactcgagga acaaaaacta
     4321 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     4381 atttccgagg aggacttata gtgagggccc gatatctcga caatcaacct ctggattaca
     4441 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4501 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4561 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     4621 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4681 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4741 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4801 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     4861 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4921 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4981 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     5041 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5101 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5161 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5221 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5281 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     5341 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5401 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5461 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     5521 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5581 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5641 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     5701 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5761 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5821 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5941 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6001 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6061 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6121 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6181 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6241 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6301 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     6361 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6421 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6481 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6541 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6601 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6661 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6721 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6781 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6841 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6901 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6961 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     7021 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7081 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     7141 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7201 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7261 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     7321 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7381 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     7441 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7501 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     7921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     8161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     9481 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     9541 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     9601 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
