LOCUS       V022808                 7895 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022808
VERSION     V022808
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             683..2542
                     /label="DTX1(NM_004416)"
                     /note="DTX1(NM_004416)"
                     /gene="DTX1"
     CDS             2582..2647
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2671..3259
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3325..3373
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3575..4003
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4017..4346
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4413..5207
                     /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    5383..5504
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5553..5569)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5577..5593
                     /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(5601..5631)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5646..5667
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5955..6543)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6714..7574)
                     /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(7575..7679)
                     /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 ggatccgcca ccatgtcacg gccaggccac ggtgggctga tgcctgtgaa
      721 tggtctgggc ttcccaccgc agaacgtggc ccgggtggtg gtgtgggagt ggctgaatga
      781 gcacagccgc tggcggccct acacggccac cgtgtgccac cacattgaga acgtgctgaa
      841 ggaggacgct cgcggttccg tggtcctggg gcaggtggac gcccagcttg tgccctacat
      901 catcgacctg cagtccatgc accagtttcg ccaggacaca ggcaccatgc ggcccgtgcg
      961 gcgcaacttc tacgacccgt cgtcggcgcc gggcaagggc atcgtgtggg agtgggagaa
     1021 cgacggcggc gcatggacgg cctacgatat ggacatctgc atcaccatcc agaacgccta
     1081 cgagaagcag cacccgtggc tcgacctctc atcgctaggc ttctgctacc tcatctactt
     1141 caacagcatg tcgcagatga accgccagac gcgccggcgc cgccgcctgc gccgccgcct
     1201 ggacctcgcc tacccgctca ccgtgggctc catccctaag tcgcagtcgt ggcccgtggg
     1261 cgccagctcg ggccagccct gctcctgcca gcagtgcctg ctggtcaaca gcacgcgcgc
     1321 cgcctccaac gccatcctgg cctcgcagcg ccgcaaggcg ccccccgcgc ccccgctgcc
     1381 gccgccgccg ccacctggag ggcctccagg cgcgcttgcc gtgcgcccca gcgccacctt
     1441 cacaggcgcc gcgctctggg cagcgcccgc cgccggcccc gccgagcccg cgccgcctcc
     1501 cggggcgccc ccacggagcc cgggcgcccc cggcggagcg cgcaccccgg ggcagaacaa
     1561 cctcaaccgg cccgggcccc agcgcaccac cagcgtgagc gcgcgcgcct ccatcccgcc
     1621 gggggtcccc gcactcccgg tgaagaactt gaatggtact gggccggtcc atccggccct
     1681 ggcagggatg accgggatac tgctgtgcgc ggccgggctg cccgtgtgcc tgacgcgggc
     1741 ccccaagccc atcctgcacc cgccgcccgt gagcaagagc gacgtgaagc ccgtgcctgg
     1801 cgtgcccggg gtgtgccgca agaccaagaa gaagcacctt aaaaagagta agaatcccga
     1861 ggatgtggtt cgaagataca tgcagaaggt gaaaaaccca cctgatgagg actgcaccat
     1921 ctgcatggag cgactggtca cagcatcagg ctacgagggc gtgcttcggc acaagggcgt
     1981 gcggcctgag ctcgtgggcc gcctgggccg ctgtggccac atgtaccacc tgctgtgcct
     2041 cgtggccatg tactccaatg gcaacaagga tggcagcctg cagtgcccca cctgcaaggc
     2101 catctacggg gagaagacgg gtacgcagcc gcctgggaag atggagttcc acctcatccc
     2161 ccactcgctg cccggcttcc ctgataccca gaccatccgc atcgtctatg acatccccac
     2221 aggcatccag ggccctgagc accccaaccc cgggaagaag ttcaccgcaa gaggattccc
     2281 tcgccactgc tatctaccca acaacgagaa aggccggaag gtgctgcggc tgctcatcac
     2341 ggcctgggag agaagactca tcttcactat cggcacgtcc aacaccacgg gcgagtcgga
     2401 caccgtggtg tggaacgaga tccaccacaa gaccgagttt ggatccaacc tcacgggcca
     2461 cggctacccg gacgctagct acctagacaa cgtgctggct gagctcacag cccagggcgt
     2521 atccgaggct gcagccaagg ctgaattctg cagatatcca gcacagtggc ggccgctcga
     2581 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2641 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2701 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2761 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2821 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2881 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2941 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3001 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3061 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3121 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3181 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3241 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3301 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3361 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3421 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3481 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3541 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3841 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3901 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3961 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4261 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4321 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4381 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4441 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4501 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4561 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4621 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4681 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4741 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4801 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4861 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4921 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4981 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5041 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5101 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5161 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5221 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5281 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5341 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5401 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5461 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5521 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5581 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5641 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5761 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6121 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6361 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7621 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7681 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7741 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7801 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7861 gcttgaccga caattgcatg aagaatctgc ttagg
//
