LOCUS       V046418                 6579 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046418
VERSION     V046418
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..1259
                     /label="ARPC4(NM_005718)"
                     /note="ARPC4(NM_005718)"
                     /gene="ARPC4"
     CDS             1266..1331
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1355..1943
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2009..2057
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2259..2687
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2701..3030
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3097..3891
                     /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    4067..4188
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4237..4253)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4261..4277
                     /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(4285..4315)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4330..4351
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4639..5227)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5398..6258)
                     /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(6259..6363)
                     /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 ccaccatgac tgccactctc cgcccctacc
      781 tgagtgccgt gcgggccaca ttgcaggctg ccctctgcct ggagaacttc tcctcccagg
      841 ttgtggaacg acacaacaag ccggaagtgg aagtcaggag tagcaaagag ctcctgttac
      901 aacctgtgac catcagcagg aatgagaagg aaaaggttct gattgagggc tccatcaact
      961 ctgtccgggt cagcattgct gtgaaacagg ctgatgagat cgagaagatt ttgtgccaca
     1021 agttcatgcg cttcatgatg atgcgagcag agaacttctt tatccttcga aggaagcctg
     1081 tggaggggta tgatatcagc tttctgatca ccaacttcca cacagagcag atgtacaaac
     1141 acaagttggt ggactttgtg atccacttca tggaggagat tgacaaggag atcagtgaga
     1201 tgaagctgtc agtcaatgcc cgtgcccgca ttgtggctga agagttcctt aagaattttg
     1261 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     1321 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     1381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1501 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1561 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1621 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1681 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     1741 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     1801 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     1861 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     1921 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     1981 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2041 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     2101 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     2161 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     2221 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2401 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2461 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2581 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     2641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2701 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2761 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2821 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2881 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2941 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3001 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3061 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3121 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3181 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3241 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3301 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3361 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3421 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3481 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3541 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3601 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3661 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3721 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3781 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3841 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3901 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3961 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4021 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4201 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4261 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4321 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4381 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4441 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4501 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4801 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     4861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5041 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5221 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5341 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5521 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5701 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5821 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6001 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6361 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6421 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6481 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6541 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
