LOCUS       V019351                 6839 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019351
VERSION     V019351
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"
     CDS             683..1519
                     /label="Zdhhc25(NM_027306)"
                     /note="Zdhhc25(NM_027306)"
                     /gene="Zdhhc25"
     CDS             1526..1591
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1615..2203
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2269..2317
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2519..2947
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2961..3290
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3357..4151
                     /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    4327..4448
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4497..4513)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4521..4537
                     /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(4545..4575)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4590..4611
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4899..5487)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5658..6518)
                     /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(6519..6623)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgtcctc gtgggtccaa aaccacggca catctgagca
      721 cagcgccggg ctagagaccc cgctgcaggc ccccaaccaa cgctgctggt tcatattgga
      781 tcccattggc atcctgtgcg ccatggccgc gtgggcgctg gtactgagcg gcggatgggt
      841 actgttccgg gacctgctta taccatctaa caacatgttg tacatagtag ccaatggtgt
      901 ggtcttccat ctgctggcct cgttggcact cgcgtcacac ctacggacta tgctgaccga
      961 cccaggctct gtgcccctgg gtaacccgcc cgggcctgac acagtgtcct actgtaccga
     1021 ctgccactct gccataccga ggacagcttg ccactgtaca gtgtgccagc gctgcatccg
     1081 caagaacgac caccactgtc cgtggattaa caactgcatt ggtgaagaca accagaagta
     1141 cttcctgctt ttcacaatgt acattggact tacctcgacc cacgttctgc tcctgttggg
     1201 catccctgtc ctgtgcagct acatgcgagg cgagtgggat tcgagcagca ccgtgtccct
     1261 acctgcccct atcctctttc tcctcctggt ggcgataatg ggcttcctct ttgccgtggt
     1321 gatgctctgc agccaaatgt gcgtcatcta ctcagacaaa acgacaacgg agctgctgta
     1381 tcagaacaca cactcagggg gcagatggtc caagtgcgca aacatgaagg ccgtctgtgg
     1441 ctcccacgtc tccctggcct ggctcagccc ttttcattct ccagaacatt acaaagtgag
     1501 tgagcaccat gatatggccc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1561 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     1621 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1681 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1741 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1801 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     1861 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     1921 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1981 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2041 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2101 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2161 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2221 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2281 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2341 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2401 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2461 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2641 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2881 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2941 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3301 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4141 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4201 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4261 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4321 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4381 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4441 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4501 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4561 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4621 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6661 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     6721 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6781 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
