LOCUS       V044873                 7875 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044873
VERSION     V044873
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..2555
                     /label="Ift74(NM_026319)"
                     /note="Ift74(NM_026319)"
                     /gene="Ift74"
     CDS             2562..2627
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2651..3239
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3305..3353
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3555..3983
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3997..4326
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4393..5187
                     /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    5363..5484
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5533..5549)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5557..5573
                     /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(5581..5611)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5626..5647
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5935..6523)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6694..7554)
                     /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(7555..7659)
                     /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 cagtaatcac aaatcttcag
      781 cccctcgccc tatttctcgg ggtggaatag gactaacagg aaggcctcct tctggaataa
      841 gacctccatc tggcaatgtt cgagtggcaa ctgcaatgcc accaacaaca gcaagaccag
      901 gttctcgtgg tggtccctta gggactggtg gagttttgtc atctcaaatc aaagttgctg
      961 atcgtcctgt gacccaacaa ggtttgagtg gaatgaagac tggcatgaaa ggtccccaga
     1021 ggcaaatttt agacaaatct tactatcttg gacttcttag gagcaaaata agtgaactta
     1081 caactgaaat taataagctt cagaaagaaa tagaaatgta caaccaagaa aattcagtgt
     1141 atttgtcata tgaaaagagg gcagagacat tagctgttga aattaaagac tttcaaggac
     1201 aactagcaga ctacaacatg ttggtagata aacttaatac caatactgaa atggaagaag
     1261 taatgagtga ttacaacatg cttaaagctc aaaatgacag agaaacacaa agtatggatg
     1321 tcatatttac tgaaagacaa gcgaaggaaa aacaaatcag aagtgtagaa gaagaagttg
     1381 aacaagaaaa gcaagcagca gatggcatta tcaaaaatat gtctcctgaa aaacaagtca
     1441 agtacataga aatgaaaacc acaaatgaga aactgttgca ggaattagat acacttcagc
     1501 aacagctaga ttctctgaac atgaaaaagg agagcctaga aactgaaata gcacactccc
     1561 aagtaaaaca agaggctgta ttgctgtatg aaaaacttta tgagttagag tcccatcgag
     1621 atcaaatgat tgcagaagac aaaagcatgg gatcgccaat ggaggaaagg gagagattgc
     1681 ttaagcaggt taaagaagat aatcaggaaa tagccagcat ggagagacag ttaacagata
     1741 taaaagaaaa aataaaccaa tttagtgaag aaattagaca acttgacatg gatttagagg
     1801 agcaccaagg tgaaatgaat caaaaataca aggagcttaa aaaaagggag gaaaatatgg
     1861 atgcttttat agagactttt gaggaaacga agaatcagga actagagcgg aaggcacaga
     1921 tagaagccag catcatcaca ctgttagagc attgcagtcg gaatataaat cgtatgaagc
     1981 agatatcctc catcaccaac caagagctga agatgatgca ggatgacctc agctttaaat
     2041 caactgagat gcagaagtca cagacaacag ctaggaattt gacctcagac agtcagcgtc
     2101 tgcagctgga tctacagaaa atggaacttc tggaaagtaa gatgacagaa gagcaacagt
     2161 ctctgaaaaa caaaattaag caaatgactg ctgacctgga gacatacagt gacctggcag
     2221 ctttaaaatc atcagctgaa gaaaagaaga agaaattgca tcaagaaaga acagtattgt
     2281 caactcacag aaatgccttt aagaaaataa tggagaagct gacctcagat tatgataccc
     2341 ttaaaaggca acttcaagac aatgagacac atgctcagct tacaaatttg gagaggaagt
     2401 ggcagcacct ggagcaaaat aacttcgtaa tgaaagagtt catagcaacc aagagtcaag
     2461 agagtgatta ccagccggtg attaaaaatg tgatgaagca gattgcagag tataataaaa
     2521 ccatcatgga tgctctacat aatgccagca gaagcggatc cgactacaaa gaccatgacg
     2581 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     2641 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2701 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2761 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2821 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2881 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2941 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3001 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3061 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3121 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3181 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3241 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3301 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3361 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3421 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3481 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3541 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3901 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3961 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4141 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4201 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4261 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4321 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4561 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4921 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5221 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5281 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5341 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5461 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5521 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5581 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5641 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5701 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5761 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5821 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5881 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5941 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6001 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6061 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6121 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6181 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6241 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6301 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6361 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6421 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6481 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6541 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6601 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6661 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6721 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6781 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6841 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6901 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6961 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7021 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7081 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7141 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7201 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7261 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7321 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7381 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7441 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7501 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7621 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7681 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7741 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7801 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7861 aagaatctgc ttagg
//
