LOCUS       V033112                 7815 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033112
VERSION     V033112
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2495
                     /label="Trafd1(NM_001163470)"
                     /note="Trafd1(NM_001163470)"
                     /gene="Trafd1"
     CDS             2502..2567
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2591..3179
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3245..3293
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3495..3923
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3937..4266
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4333..5127
                     /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    5303..5424
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5473..5489)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5497..5513
                     /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(5521..5551)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5566..5587
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5875..6463)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6634..7494)
                     /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(7495..7599)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cgagtttcga gatgaccagg
      781 cttctaggct gtgtgacaac tgcaagaagg aaattcctgt atttaatttt accatccatg
      841 aaatccactg tcaaaggaac attggtgtgt gccctgtctg caaggaaccg ttccccaaat
      901 ctgacatgga cattcacatg gctgcagagc actgtcaggt gacctgtaaa tgcaacaaga
      961 agttggagaa gaggcagtta aagcagcatg cggagacaga gtgtcccctg cggctcgccg
     1021 tctgccagca ctgtgatctg gagctttctg ttgtcaagtt gaaggagcat gaggattact
     1081 gtggagcccg gacagagctg tgtggcagct gtgggcgcaa cgtgcttgtg aaggagctga
     1141 agactcaccc cgaagtctgt gggagagtgg aggaggaaaa gagaacggag gctgccatcc
     1201 ctccggaggc ttacgacgag ccctggagtc aggacagaat ctggatcgca tcccagctcc
     1261 tcagacaaat cgaggctctg gacccgccca tgaggctccc tggaaggccc ctgcaagcct
     1321 ttgaggcaga ccccttctac agtaggacta ccagccagag gagcatggca gcccagtttc
     1381 cagttcaaaa taatcttttt gaagaacaag aaaggcagga aaggaacaga agccggcagt
     1441 ccccaaagga cagcgctgag aataacgcac acttggactt catgttggcc ttgagtctgc
     1501 agaatgaggg acaggccacc agcatggtag agcagggctt ctgggagtct gtgcctgagg
     1561 ctgatccggc tcgtgctggg cccacatctc taggtgacat aaagggtgct gctgacgaga
     1621 ttctgctgcc gtgtgagttc tgtgaggagc tgtacccaga ggaactgctc attgaccatc
     1681 agacaagctg caacccttct catgccttac gttcactcaa tacgggcagc tcttccatca
     1741 ggggtgtgga agatcctggt accatcttcc agaactttct acaacaagca acaagtaacc
     1801 agtttgacac tttaatgggc ctgagcagtt cagctgctgt ggaagacagc atcatcatcc
     1861 cctgtgagtt ctgtggggtg cagctggaag aggaggtgct gttctaccat caggaccagt
     1921 gtgaccaacg cccagccaca gcaaaccacc gtgcagtgga gggcatccca gcccaggatt
     1981 cgcagccaga aaacacttca gcagagctgt ccaggaggcg ggtcaaacac cagggagacc
     2041 tgtcatctgg ttacatggat gatgtcaagc cggaatcagt gaaaggcccc acctactcga
     2101 tgtctcctaa caggaccatg aacaatgtgg cttcctgcaa ccgactgttg aacttaccgt
     2161 cagggcccag gtctgactgc cagcgtagcc ctcccggtgt gctgaaactc aacaactctg
     2221 atagccagga catccgtggg cagatgcggg gcagccagaa tgggcccata gcatccgggc
     2281 acgctccagt gatccactct attcaaaatc tctatccaga aaactttgcg ccctcttttc
     2341 ctcatggatc ccctgggagg tacggagctg gtgggaggag tgaaggtggt cggagctctc
     2401 gggtcagccc tgcagctgct ggctaccaca gcagagcagc aaaggccaag cctccaaagc
     2461 agcagggagc aggcgatgcg gaggaggaag aggagggatc cgactacaaa gaccatgacg
     2521 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     2581 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2641 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2701 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2761 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2821 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2881 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2941 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3001 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3061 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3121 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3181 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3241 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3301 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3361 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3421 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3481 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3541 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3601 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3661 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3721 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3781 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3841 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3901 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4021 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4081 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4141 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4201 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4261 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4321 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4501 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4861 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5101 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5281 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5341 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5401 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7621 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7681 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7741 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7801 aagaatctgc ttagg
//
