LOCUS       V035652                 9312 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035652
VERSION     V035652
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..3992
                     /label="ZRANB3(NM_032143)"
                     /note="ZRANB3(NM_032143)"
                     /gene="ZRANB3"
     CDS             3999..4064
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4088..4676
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4742..4790
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4992..5420
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5434..5763
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5830..6624
                     /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    6800..6921
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6970..6986)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6994..7010
                     /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(7018..7048)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7063..7084
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7372..7960)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8131..8991)
                     /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(8992..9096)
                     /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 ccaccatgcc tagggttcat aacataaaaa
      781 agtctcttac acctcacatt tcttgtgtga caaatgaatc tgataatctg ctggattttt
      841 tgcctgacag actaagagca aagctacttc cattccagaa agatggcatc atttttgccc
      901 tcaaaagaaa tggcaggtgt atggtggctg atgaaatggg tctaggaaag acaatccagg
      961 caattggaat tacttacttc tataaagagg aatggcctct gttaatagtg gtcccttcgt
     1021 ctctgaggta cccttggaca gaagaaattg aaaaatggat cccagagcta agtccagaag
     1081 aaatcaatgt tattcagaat aaaactgatg ttaggagaat gtcgaccagt aaagtgacag
     1141 ttctgggtta tggtctctta accgcagatg caaagacttt gatagatgca ctgaataatc
     1201 agaacttcaa agtagttata gtggatgaat cacactacat gaaatccaga aatgcaactc
     1261 gcagcaggat tttattgcca atagtacaga aagccagacg agccattctt cttacaggaa
     1321 caccagcttt aggaaggcct gaagagcttt ttatgcagat tgaagctctc tttccacaaa
     1381 aatttggaag atggaccgac tatgcaaaaa gatactgtaa tgcacacatc agatattttg
     1441 gtaaaagacc tcagtgggat tgtagagggg catcaaatct taatgaactt caccagctat
     1501 taagtgacat aatgattaga agattaaaga ctgaagtttt aacccagcta ccccctaaag
     1561 tcagacagcg tattccattt gatcttccat cagcagctgc caaggaattg aataccagct
     1621 ttgaagagtg ggaaaaaata atgagaactc caaattcagg tgccatggag acagtcatgg
     1681 ggttgataac tcgcatgttt aaacaaactg ctattgccaa ggcaggtgct gtaaaggatt
     1741 atattaagat gatgcttcag aatgattcgc ttaaatttct ggtttttgct caccatttaa
     1801 gcatgctcca agcttgcaca gaagcagtca tcgaaaataa gactcgttac attaggatag
     1861 atggaagtgt ttcatcttca gaaagaatac atctggttaa tcagtttcaa aaggatcctg
     1921 acactcgcgt ggctatccta agcattcagg ctgctggcca gggattaaca tttactgcag
     1981 caagtcatgt tgtatttgct gagttgtact gggaccctgg acatataaaa caagcagaag
     2041 accgagctca cagaattggc cagtgcagtt ctgtgaatat tcactacctt attgcaaatg
     2101 gaaccctaga cacccttatg tggggaatgt tgaatcgcaa ggctcaagtt acagggagca
     2161 cactgaacgg taggaaagaa aaaattcagg ctgaggaagg tgataaggaa aaatgggatt
     2221 tcctgcagtt tgctgaagct tggactccaa atgacagttc tgaagagtta aggaaggaag
     2281 ctttgttcac tcacttcgaa aaagaaaaac agcatgatat tcgatcattt tttgtaccac
     2341 aacctaaaaa aagacagttg atgacctcct gtgacgaatc aaagagattc cgggaggaaa
     2401 atactgtagt gtcatcagac cctacaaaaa cagctgcaag agatatcatc gattatgaaa
     2461 gtgatgttga acctgaaacg aaaagattga aattggctgc ctcggaagac cactgcagtc
     2521 cgtcggaaga gacaccatcc cagtccaagc aaatccgaac tccactcgtg gaaagtgtac
     2581 aggaggccaa ggcccagtta accaccccag cctttcctgt agagggctgg caatgtagtc
     2641 tctgcaccta tatcaataat tcagagttac cttattgtga aatgtgtgag actcctcaag
     2701 gcagtgctgt tatgcaaata gatagcctca accatatcca ggataaaaac gagaaggatg
     2761 attctcagaa agacacctcc aaaaaggttc aaactatctc agactgtgaa aaacaagccc
     2821 ttgcacagtc agaacctggc cagttggctg acagcaagga agaaacacca aaaattgaga
     2881 aagaagacgg acttacatcc cagccaggta atgaacagtg gaagagttca gacactttgc
     2941 cagtgtatga caccttaatg ttctgtgcaa gtaggaatac tgaccggatt cacatctata
     3001 ctaaggatgg aaaacagatg agctgtaatt tcattcctct ggatataaaa ttagaccttt
     3061 gggaagattt accagcaagc tttcagctga aacaatatcg ctcactgatt ttgagatttg
     3121 ttcgagaatg gagtagtcta actgccatga agcaaaggat aatcaggaaa agtggacagc
     3181 tattctgtag cccaattctt gctttggaag agatcacaaa gcaacaaacc aaacaaaatt
     3241 gcaccaaaag atacataacc aaagaagatg ttgccgtagc ctcaatggac aaagtgaaga
     3301 atgttggggg ccatgtccgt ctgatcacaa aggagtccag gccacgggat cctttcacaa
     3361 aaaaacttct tgaagatgga gcctgtgtcc catttctaaa tccatacaca gtccaggcag
     3421 atctcactgt gaagccctct acatccaaag gctatttgca agctgtggat aatgaaggaa
     3481 atccactttg ccttcgctgt cagcaaccca cttgccagac taagcaagca tgtaaagcga
     3541 actcttggga ttcacggttt tgctctctga aatgtcagga agagttttgg attcgatcta
     3601 ataacagtta cctgagagcc aaagtatttg aaactgaaca tggtgtgtgt cagctctgta
     3661 atgtgaacgc acaagaactc tttttacgtc tgagagatgc ccctaaaagt cagaggaaga
     3721 atcttctgta tgctacctgg acttcaaagc tcccattaga acagctaaat gaaatgataa
     3781 gaaacccagg ggaaggacat ttctggcagg tggatcacat caagccagtg tatgggggag
     3841 gaggacagtg ttccctggac aacctgcaga ctctctgcac agtctgtcac aaagagagaa
     3901 ctgccagaca agctaaggaa agaagccagg tgagaagaca atctctagca tcaaagcatg
     3961 gatcagacat cacacgattt ttggtaaaga agggatccga ctacaaagac catgacggtg
     4021 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     4081 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4141 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4201 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4261 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4321 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4381 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4441 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4501 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4561 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4621 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4681 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4741 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4801 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4861 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4921 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4981 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5041 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5101 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     5161 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5221 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5281 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5341 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     5401 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5461 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5521 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5581 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5641 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5701 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5761 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5821 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5881 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5941 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6001 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6061 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6121 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6181 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6241 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6301 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6361 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6421 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6481 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6541 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6601 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6661 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6721 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6781 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6841 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6901 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6961 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7021 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7081 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7141 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7261 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7561 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7801 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9121 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     9181 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9241 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     9301 aatctgctta gg
//
