LOCUS       V018921                 9293 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018921
VERSION     V018921
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..3973
                     /label="LIFR(NM_001127671)"
                     /note="LIFR(NM_001127671)"
                     /gene="LIFR"
     CDS             3980..4045
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4069..4657
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4723..4771
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4973..5401
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5415..5744
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5811..6605
                     /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    6781..6902
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6951..6967)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6975..6991
                     /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(6999..7029)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7044..7065
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7353..7941)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8112..8972)
                     /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(8973..9077)
                     /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 ccatgatgga tatttacgta tgtttgaaac gaccatcctg
      721 gatggtggac aataaaagaa tgaggactgc ttcaaatttc cagtggctgt tatcaacatt
      781 tattcttcta tatctaatga atcaagtaaa tagccagaaa aagggggctc ctcatgattt
      841 gaagtgtgta actaacaatt tgcaagtgtg gaactgttct tggaaagcac cctctggaac
      901 aggccgtggt actgattatg aagtttgcat tgaaaacagg tcccgttctt gttatcagtt
      961 ggagaaaacc agtattaaaa ttccagctct ttcacatggt gattatgaaa taacaataaa
     1021 ttctctacat gattttggaa gttctacaag taaattcaca ctaaatgaac aaaacgtttc
     1081 cttaattcca gatactccag agatcttgaa tttgtctgct gatttctcaa cctctacatt
     1141 atacctaaag tggaacgaca ggggttcagt ttttccacac cgctcaaatg ttatctggga
     1201 aattaaagtt ctacgtaaag agagtatgga gctcgtaaaa ttagtgaccc acaacacaac
     1261 tctgaatggc aaagatacac ttcatcactg gagttgggcc tcagatatgc ccttggaatg
     1321 tgccattcat tttgtggaaa ttagatgcta cattgacaat cttcattttt ctggtctcga
     1381 agagtggagt gactggagcc ctgtgaagaa catttcttgg atacctgatt ctcagactaa
     1441 ggtttttcct caagataaag tgatacttgt aggctcagac ataacatttt gttgtgtgag
     1501 tcaagaaaaa gtgttatcag cactgattgg ccatacaaac tgccccttga tccatcttga
     1561 tggggaaaat gttgcaatca agattcgtaa tatttctgtt tctgcaagta gtggaacaaa
     1621 tgtagttttt acaaccgaag ataacatatt tggaaccgtt atttttgctg gatatccacc
     1681 agatactcct caacaactga attgtgagac acatgattta aaagaaatta tatgtagttg
     1741 gaatccagga agggtgacag cgttggtggg cccacgtgct acaagctaca ctttagttga
     1801 aagtttttca ggaaaatatg ttagacttaa aagagctgaa gcacctacaa acgaaagcta
     1861 tcaattatta tttcaaatgc ttccaaatca agaaatatat aattttactt tgaatgctca
     1921 caatccgctg ggtcgatcac aatcaacaat tttagttaat ataactgaaa aagtttatcc
     1981 ccatactcct acttcattca aagtgaagga tattaattca acagctgtta aactttcttg
     2041 gcatttacca ggcaactttg caaagattaa ttttttatgt gaaattgaaa ttaagaaatc
     2101 taattcagta caagagcagc ggaatgtcac aatcaaagga gtagaaaatt caagttatct
     2161 tgttgctctg gacaagttaa atccatacac tctatatact tttcggattc gttgttctac
     2221 tgaaactttc tggaaatgga gcaaatggag caataaaaaa caacatttaa caacagaagc
     2281 cagtccttca aaggggcctg atacttggag agagtggagt tctgatggaa aaaatttaat
     2341 aatctattgg aagcctttac ccattaatga agctaatgga aaaatacttt cctacaatgt
     2401 atcgtgttca tcagatgagg aaacacagtc cctttctgaa atccctgatc ctcagcacaa
     2461 agcagagata cgacttgata agaatgacta catcatcagc gtagtggcta aaaattctgt
     2521 gggctcatca ccaccttcca aaatagcgag tatggaaatt ccaaatgatg atctcaaaat
     2581 agaacaagtt gttgggatgg gaaaggggat tctcctcacc tggcattacg accccaacat
     2641 gacttgcgac tacgtcatta agtggtgtaa ctcgtctcgg tcggaaccat gccttatgga
     2701 ctggagaaaa gttccctcaa acagcactga aactgtaata gaatctgatg agtttcgacc
     2761 aggtataaga tataattttt tcctgtatgg atgcagaaat caaggatatc aattattacg
     2821 ctccatgatt ggatatatag aagaattggc tcccattgtt gcaccaaatt ttactgttga
     2881 ggatacttct gcagattcga tattagtaaa atgggaagac attcctgtgg aagaacttag
     2941 aggcttttta agaggatatt tgttttactt tggaaaagga gaaagagaca catctaagat
     3001 gagggtttta gaatcaggtc gttctgacat aaaagttaag aatattactg acatatccca
     3061 gaagacactg agaattgctg atcttcaagg taaaacaagt taccacctgg tcttgcgagc
     3121 ctatacagat ggtggagtgg gcccggagaa gagtatgtat gtggtgacaa aggaaaattc
     3181 tgtgggatta attattgcca ttctcatccc agtggcagtg gctgtcattg ttggagtggt
     3241 gacaagtatc ctttgctatc ggaaacgaga atggattaaa gaaaccttct accctgatat
     3301 tccaaatcca gaaaactgta aagcattaca gtttcaaaag agtgtctgtg agggaagcag
     3361 tgctcttaaa acattggaaa tgaatccttg taccccaaat aatgttgagg ttctggaaac
     3421 tcgatcagca tttcctaaaa tagaagatac agaaataatt tccccagtag ctgagcgtcc
     3481 tgaagatcgc tctgatgcag agcctgaaaa ccatgtggtt gtgtcctatt gtccacccat
     3541 cattgaggaa gaaataccaa acccagccgc agatgaagct ggagggactg cacaggttat
     3601 ttacattgat gttcagtcga tgtatcagcc tcaagcaaaa ccagaagaag aacaagaaaa
     3661 tgaccctgta ggaggggcag gctataagcc acagatgcac ctccccatta attctactgt
     3721 ggaagatata gctgcagaag aggacttaga taaaactgcg ggttacagac ctcaggccaa
     3781 tgtaaataca tggaatttag tgtctccaga ctctcctaga tccatagaca gcaacagtga
     3841 gattgtctca tttggaagtc catgctccat taattcccga caatttttga ttcctcctaa
     3901 agatgaagac tctcctaaat ctaatggagg agggtggtcc tttacaaact tttttcagaa
     3961 caaaccaaac gatctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     4021 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     4081 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4141 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4201 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4261 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4321 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4381 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4441 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     4501 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4561 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4621 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     4681 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4741 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4801 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4861 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4921 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     4981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5101 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     5161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5341 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5401 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5461 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5701 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5761 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5821 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5881 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5941 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     6001 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6061 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6121 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6181 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6241 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6301 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     6361 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6421 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6481 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6541 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6601 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     6661 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6721 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6781 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6901 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     7561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7981 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9121 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     9181 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     9241 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
