LOCUS       V018516                 7384 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018516
VERSION     V018516
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..2863
                     /label="KRT1(NM_006121)"
                     /note="KRT1(NM_006121)"
                     /gene="KRT1"
     CDS             2885..2950
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2979..3203
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3249..3677
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3691..4020
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4087..4881
                     /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    5057..5178
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5227..5243)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5251..5267
                     /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(5275..5305)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5320..5341
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6388..7248)
                     /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(7249..7353)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgagtcga cagtttagtt ccaggtctgg
      961 gtaccgaagt ggagggggct tcagctctgg ctctgctggg atcatcaact accagcgcag
     1021 gaccaccagc agctccacac gccgcagtgg aggaggtggt gggagatttt caagctgtgg
     1081 tggtggtggt ggtagctttg gtgctggtgg tggatttgga agtcggagtc ttgttaacct
     1141 tggtggcagt aaaagcatct ccataagtgt ggctagagga ggtggacgtg gtagtggctt
     1201 tggtggtggt tatggtggtg gtggctttgg tggtggtggc tttggtggtg gtggctttgg
     1261 tggaggtggc attgggggtg gtggctttgg tggttttggc agtggtggtg gtggttttgg
     1321 tggaggtggc tttgggggtg gtggatatgg gggtggttat ggtcctgtct gccctcctgg
     1381 tggcatacaa gaagtcacta tcaaccagag ccttcttcag cccctcaatg tggagattga
     1441 ccctgagatc caaaaggtga agtctcgaga aagggagcaa atcaagtcac tcaacaacca
     1501 atttgcctcc ttcattgaca aggtgaggtt cctggagcag cagaaccagg tactgcaaac
     1561 aaaatgggag ctgctgcagc aggtagatac ctccactaga acccataatt tagagcccta
     1621 ctttgagtca ttcatcaaca atctccgaag gagagtggac caactgaaga gtgatcaatc
     1681 tcggttggat tcggaactga agaacatgca ggacatggtg gaggattacc ggaacaagta
     1741 tgaggatgaa atcaacaagc ggacaaatgc agagaatgaa tttgtgacca tcaagaagga
     1801 tgtggatggt gcttatatga ccaaggtgga ccttcaggcc aaacttgaca acttgcagca
     1861 ggaaattgat ttccttacag cactctacca agcagagttg tctcagatgc agactcaaat
     1921 cagtgaaact aatgtcatcc tctctatgga caacaaccgc agtctcgacc tggacagcat
     1981 cattgctgag gtcaaggccc agtacgagga tatagcccag aagagcaaag ctgaggccga
     2041 gtccttgtac cagagcaagt atgaagagct gcagatcact gctggcagac atggggatag
     2101 tgtgagaaat tcaaagatag aaatttctga gctgaatcgt gtgatccaga gacttagatc
     2161 tgaaatcgac aatgtcaaga agcagatctc caacttgcag cagtccatca gtgatgcaga
     2221 gcagcgtggc gagaatgccc tcaaggatgc caagaacaag ctgaatgacc tggaggatgc
     2281 cctgcagcag gccaaggaag acctggcccg cctgctgcgc gactaccagg agctgatgaa
     2341 cacaaagctg gccctggatc tggagattgc cacctacagg accctcctgg agggagaaga
     2401 aagcaggatg tctggagaat gtgccccgaa cgtgagtgtg tctgtgagca caagccacac
     2461 caccatcagt ggaggtggca gccgaggagg tggcggcggt ggctacggct ctggaggtag
     2521 cagctatggc tccggaggtg gtagctatgg ttctggaggt ggcggcggcg gcggccgtgg
     2581 cagctatggc tccggaggta gcagctacgg ctccggaggt ggcagctatg gctctggagg
     2641 tggcggcggc ggccatggca gctacggctc cggaagcagc agtgggggct acagaggtgg
     2701 ctctggaggc ggcggcggcg gcagctctgg cggccggggc tctggcggcg ggagctctgg
     2761 aggctccata ggaggccggg gatccagctc tgggggtgtc aagtcctctg gtggcagttc
     2821 cagcgtgaag tttgtttcta ccacttattc cggagtaacc agactcgagt ctagagggcc
     2881 cttcgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2941 cgatgacaag tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3001 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3061 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3121 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3181 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3241 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3301 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3361 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3421 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3481 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3541 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3601 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3661 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3721 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3781 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3841 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3901 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3961 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4021 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4081 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4141 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4201 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4261 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4321 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4381 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4441 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4501 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4561 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4621 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4681 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4741 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4801 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4861 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4921 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4981 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5041 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5101 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5161 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5281 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7381 cgtc
//
