LOCUS       V042161                 7752 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042161
VERSION     V042161
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..2432
                     /label="Dennd6a(NM_145969)"
                     /note="Dennd6a(NM_145969)"
                     /gene="Dennd6a"
     CDS             2439..2504
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2528..3116
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3182..3230
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3432..3860
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3874..4203
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4270..5064
                     /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    5240..5361
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5410..5426)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5434..5450
                     /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(5458..5488)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5503..5524
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5812..6400)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6571..7431)
                     /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(7432..7536)
                     /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 tctgccgggc ccggcggtct
      781 ttgggcccgg ttcccgcggg tcgttggacg aggccggggc tgaggggcgc gaggcggcgg
      841 cccttgcggc ggcgggagtc gcgctggagg acgaggagga ggacgatggc cgccggggcc
      901 tgctgcgctg ggacggcttc tcggcctggc tacactgcgt gtgtgtggtg ggcttcgacc
      961 tggagctggg ccaggcggtg gaggtgatat atcctcagca ttccaaactt actgataaag
     1021 aaaaaaccaa tatttgctac ttgtcttttc cagactcaaa ttcaggttgt cttggagata
     1081 cccagttttg ttttagattc cgacagtctt ctgggagaag ggtgtcacta cactgtcttc
     1141 ttgatgaatt tgacaaagat ttaccagttt acttaaagaa agatcctgca tatttttatg
     1201 gatatgtgta ttttcgacaa gttcgagata aaactcttaa aagaggctac tttcaaaagt
     1261 cattggtttt gatcagcaaa ctaccttata ttcatttttt tcacaccgta ctcaaacaga
     1321 tagcaccaga gtattttgaa aaaaatgaac cttacttgga agcagcttgt aatgatgttg
     1381 accgatggcc tgccccagtg ccagggaaaa cgctgcacct gcctatcatg ggactagtaa
     1441 tgaaggttcg gattcccacg tgtcatgaca agcctgggac cacgcagatg gtgcagttaa
     1501 ctcagcaggc agatacacac acatctatta ttttgcctac tgttcacgag gtggatcttt
     1561 tcaggtgttt ctgcccagtt tttcttcaca gtcagatgct ctgggagttg gtgctcttgg
     1621 gagagcccct ggtggtcatg gcgccatcgc cgtcagaatc ttcagaaact gtactggctc
     1681 ttgttaactg tatctctcca ttaaagtact ttagtgattt tcggccttac ttcacgattc
     1741 atgatagtga attcaaagaa tatactaccc gtactcaagc tccgccctca gtcatcttag
     1801 gagtaaccaa cccctttttt gctaaaacac tacagcactg gccacacatt attcgaatag
     1861 gagatcttaa acctgcaggt gaaattccta agcaagttaa agtgaaaaag ctgaagaacc
     1921 taaaaactct ggattctaaa cctggagttt atacttctta caagccatat ctaaacagag
     1981 atgaggagat cataaaacaa cttcagaagg gtatacagca gaagcgtcct tctgaggccc
     2041 aaagtgttat tctccggcgc tattttttgg aactaacaca aagtttcatc attccattag
     2101 aaagatatgt ggcaagcttg atgcctttgc agaaaagtat ttctccttgg aagagtccac
     2161 cccagttacg gcagttcctt ccagaagaat ttatgaaaac acttgaaaaa acagggcctc
     2221 agctcacctc tggaataaag ggcgactgga ttggacttta ccggcagttt ctaaagtctc
     2281 caaattttga tggctggttc aagacccggc ggaaagaaat gactcaaaaa ttggaggcac
     2341 ttcatctaga agctctttgt gaagaggtca gaaaaaagtg tattagtaca tgcaagtgtt
     2401 cagagtacaa aagtattctt agaggcttag agggatccga ctacaaagac catgacggtg
     2461 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2521 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3121 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3181 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3241 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3301 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3361 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3421 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3781 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3841 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4081 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4141 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4201 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4261 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4321 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4381 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4441 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4501 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4561 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4621 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4681 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4741 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4801 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4861 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4921 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4981 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5041 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5101 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5401 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5461 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5521 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5581 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6001 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6241 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6961 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7561 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7621 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7681 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7741 aatctgctta gg
//
