LOCUS       V027616                 8803 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027616
VERSION     V027616
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             706..3462
                     /label="LIG1(NM_000234)"
                     /note="LIG1(NM_000234)"
                     /gene="LIG1"
     CDS             3469..3495
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3499..3525
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3529..3555
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3579..4167
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4233..4281
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4483..4911
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4925..5254
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5321..6115
                     /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    6291..6412
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6461..6477)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6485..6501
                     /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(6509..6539)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6554..6575
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7622..8482)
                     /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(8483..8587)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgca gcgaagtatc
      721 atgtcatttt tccaccccaa gaaagagggt aaagcaaaga agcctgagaa ggaggcatcc
      781 aatagcagca gagagacgga gccccctcca aaggcggcac tgaaggagtg gaatggagtg
      841 gtgtccgaga gtgactctcc ggtgaagagg ccagggagga aggcggcccg ggtcctgggc
      901 agcgaagggg aagaggagga tgaagccctt agccctgcta aaggccagaa gcctgccctg
      961 gactgctcac aggtctcccc gccccgtcct gccacatctc ctgagaacaa tgcttccctc
     1021 tctgacacct ctcccatgga cagttcccca tcagggattc cgaagcgtcg cacagctcgg
     1081 aagcagctcc cgaaacggac cattcaggaa gtcctggaag agcagagtga ggacgaggac
     1141 agagaagcca agaggaagaa ggaggaggaa gaagaggaga ccccgaaaga aagcctcaca
     1201 gaggctgaag tggccacaga gaaggaagga gaagacgggg accagcccac cacgcctccc
     1261 aagcccctaa agacctccaa agcagagacc ccgacggaaa gcgtttcaga gcctgaggtg
     1321 gccacgaagc aggaactgca ggaggaggaa gagcagacca agcctccccg cagagctccc
     1381 aagacgctca gcagcttctt caccccccgg aagccagcag tcaaaaaaga agtgaaggaa
     1441 gaggagccag gggctccagg aaaggaggga gctgctgagg gacccctgga tccatctggt
     1501 tacaatcctg ccaagaacaa ctatcatccc gtggaagatg cctgctggaa accgggccag
     1561 aaggttcctt acctggctgt ggcccggacg tttgagaaga tcgaggaggt gtctgctcgg
     1621 ctccggatgg tggagacgct gagcaacttg ctgcgctccg tggtggccct gtcgcctcca
     1681 gacctcctcc ctgtcctcta cctcagcctc aaccaccttg ggccacccca gcagggcctg
     1741 gagcttggcg tgggtgatgg tgtccttctc aaggcagtgg cccaggccac aggtcggcag
     1801 ctggagtccg tccgggctga ggcagccgag aaaggcgacg tggggctggt ggccgagaac
     1861 agccgcagca cccagaggct catgctgcca ccacctccgc tcactgcctc cggggtcttc
     1921 agcaagttcc gcgacatcgc caggctcact ggcagtgctt ccacagccaa gaagatagac
     1981 atcatcaaag gcctctttgt ggcctgccgc cactcagaag cccggttcat cgctaggtcc
     2041 ctgagcggac ggctgcgcct tgggctggca gagcagtcgg tgctggctgc cctctcccag
     2101 gcagtgagcc tcacgccccc gggccaagaa ttcccaccag ccatggtgga tgctgggaag
     2161 ggcaagacag cagaggccag aaagacgtgg ctggaggagc aaggcatgat cctgaagcag
     2221 acgttctgcg aggttcccga cctggaccga attatccccg tgctgctgga gcacggcctg
     2281 gaacgtctcc cggagcactg caagctgagc ccagggattc ccctgaaacc aatgttggcc
     2341 catcccaccc ggggcatcag cgaggtcctg aaacgctttg aggaggcagc tttcacctgc
     2401 gaatacaaat atgacgggca gagggcacag atccacgccc tggaaggcgg ggaggtgaag
     2461 atcttcagca ggaatcagga agacaacact gggaagtacc cggacatcat cagccgcatc
     2521 cccaagatta aactcccatc ggtcacatcc ttcatcctgg acaccgaagc cgtggcttgg
     2581 gaccgggaaa agaagcagat ccagccattc caagtgctca ccacccgcaa acgcaaggag
     2641 gtggatgcgt ctgagatcca ggtgcaggtg tgtttgtacg ccttcgacct catctacctc
     2701 aatggagagt ccctggtacg tgagcccctt tcccggcgcc ggcagctgct ccgggagaac
     2761 tttgtggaga cagagggcga gtttgtcttc gccacctccc tggacaccaa ggacatcgag
     2821 cagatcgccg agttcctgga gcagtcagtg aaagactcct gcgaggggct gatggtgaag
     2881 accctggatg ttgatgccac ctacgagatc gccaagagat cgcacaactg gctcaagctg
     2941 aagaaggact accttgatgg cgtgggtgac accctggacc tggtggtgat cggcgcctac
     3001 ctgggccggg ggaagcgggc cggccggtac gggggcttcc tgctggcctc ctacgacgag
     3061 gacagtgagg agctgcaggc catatgcaag cttggaactg gcttcagtga tgaggagctg
     3121 gaggagcatc accagagcct caaggcgctg gtgctgccca gcccacgccc ttacgtgcgg
     3181 atagatggcg ctgtgattcc cgaccactgg ctggacccca gcgctgtgtg ggaggtgaag
     3241 tgcgctgacc tctccctctc tcccatctac cctgctgcgc ggggcctggt ggatagtgac
     3301 aagggcatct cccttcgctt ccctcggttt attcgagtcc gtgaagacaa gcagccggag
     3361 caggccacca ccagtgctca ggtggcctgt ttgtaccgga agcaaagtca gattcagaac
     3421 caacaaggcg aggactcagg ctctgaccct gaagatacct acctcgagta cccatacgac
     3481 gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta tccttatgat
     3541 gtcccagact acgcatagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa
     3601 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3661 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3721 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3781 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3841 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3901 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3961 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     4021 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4081 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4141 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     4201 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4261 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4321 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4381 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4441 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     4501 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4561 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4621 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     4681 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4741 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4801 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     4861 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4921 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4981 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5101 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5161 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5221 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5281 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5341 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5401 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5461 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5521 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5581 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5641 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5701 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5761 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5821 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5881 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5941 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     6001 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     6061 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     6121 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6181 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6241 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6421 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     8641 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     8701 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     8761 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
