LOCUS       V020688                 8513 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020688
VERSION     V020688
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..3193
                     /label="Hif1a(NM_001313919)"
                     /note="Hif1a(NM_001313919)"
                     /gene="Hif1a"
     CDS             3200..3265
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3289..3877
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3943..3991
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4193..4621
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4635..4964
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5031..5825
                     /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    6001..6122
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6171..6187)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6195..6211
                     /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(6219..6249)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6264..6285
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6573..7161)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7332..8192)
                     /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(8193..8297)
                     /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 ccatggaggg cgccggcggc gagaacgaga agaaaaatag
      721 gatgagttct gaacgtcgaa aagaaaagtc tagagatgca gcaagatctc ggcgaagcaa
      781 agagtctgaa gttttttatg agcttgctca tcagttgcca cttccccaca atgtgagctc
      841 acatcttgat aaagcttctg ttatgaggct caccatcagt tatttacgtg tgagaaaact
      901 tctggatgcc ggtggtctag acagtgaaga tgagatgaag gcacagatgg actgttttta
      961 tctgaaagcc ctagatggct ttgtgatggt gctaacagat gacggcgaca tggtttacat
     1021 ttctgataac gtgaacaaat acatggggtt aactcagttt gaactaactg gacacagtgt
     1081 gtttgatttt actcatccat gtgaccatga ggaaatgaga gaaatgctta cacacagaaa
     1141 tggcccagtg agaaaaggga aagaactaaa cacacagcgg agcttttttc tcagaatgaa
     1201 gtgcacccta acaagccggg ggaggacgat gaacatcaag tcagcaacgt ggaaggtgct
     1261 tcactgcacg ggccatattc atgtctatga taccaacagt aaccaacctc agtgtgggta
     1321 caagaaacca cccatgacgt gcttggtgct gatttgtgaa cccattcctc atccgtcaaa
     1381 tattgaaatt cctttagata gcaagacatt tctcagtcga cacagcctcg atatgaaatt
     1441 ttcttactgt gatgaaagaa ttactgagtt gatgggttat gagccggaag aacttttggg
     1501 ccgctcaatt tatgaatatt atcatgcttt ggattctgat catctgacca aaactcacca
     1561 tgatatgttt actaaaggac aagtcaccac aggacagtac aggatgcttg ccaaaagagg
     1621 tggatatgtc tgggttgaaa ctcaagcaac tgtcatatat aatacgaaga actcccagcc
     1681 acagtgcatt gtgtgtgtga attatgttgt aagtggtatt attcagcacg acttgatttt
     1741 ctcccttcaa caaacagaat ctgtgctcaa accagttgaa tcttcagata tgaagatgac
     1801 tcagctgttc accaaagttg aatcagagga tacaagctgc ctttttgata agcttaagaa
     1861 ggagcctgat gctctcactc tgctggctcc agctgccggc gacaccatca tctctctgga
     1921 ttttggcagc gatgacacag aaactgaaga tcaacaactt gaagatgttc cattatataa
     1981 tgatgtaatg tttccctctt ctaatgaaaa attaaatata aacctggcaa tgtctccttt
     2041 accttcatcg gaaactccaa agccacttcg aagtagtgct gatcctgcac tgaatcaaga
     2101 ggttgcatta aaattagaat caagtccaga gtcactggga ctttctttta ccatgcccca
     2161 gattcaagat cagccagcaa gtccttctga tggaagcact agacaaagtt cacctgagag
     2221 acttcttcag gaaaacgtaa acactcctaa cttttcccag cctaacagtc ccagtgaata
     2281 ttgctttgat gtggatagcg atatggtcaa tgtattcaag ttggaactgg tggaaaaact
     2341 gtttgctgaa gacacagagg caaagaatcc attttcaact caggacactg atttagattt
     2401 ggagatgctg gctccctata tcccaatgga tgatgatttc cagttacgtt cctttgatca
     2461 gttgtcacca ttagagagca attctccaag ccctccaagt atgagcacag ttactgggtt
     2521 ccagcagacc cagttacaga aacctaccat cactgccact gccaccacaa ctgccaccac
     2581 tgatgaatca aaaacagaga cgaaggacaa taaagaagat attaaaatac tgattgcatc
     2641 tccatcttct acccaagtac ctcaagaaac gaccactgct aaggcatcag catacagtgg
     2701 cactcacagt cggacagcct caccagacag agcaggaaag agagtcatag aacagacaga
     2761 caaagctcat ccaaggagcc ttaacctgtc tgccactttg aatcaaagaa atactgttcc
     2821 tgaggaagaa ttaaacccaa agacaatagc ttcgcagaat gctcagagga agcgaaaaat
     2881 ggaacatgat ggctcccttt ttcaagcagc aggaattgga acattattgc agcaaccagg
     2941 tgactgtgca cctactatgt cactttcctg gaaacgagtg aaaggattca tatctagtga
     3001 acagaatgga acggagcaaa agactattat tttaataccc tccgatttag catgcagact
     3061 gctggggcag tcaatggatg agagtggatt accacagctg accagttacg attgtgaagt
     3121 taatgctccc atacaaggca gcagaaacct actgcagggt gaagaattac tcagagcttt
     3181 ggatcaagtt aacctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     3241 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     3901 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3961 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4021 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4081 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4141 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     4201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4261 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4321 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4381 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4501 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4561 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4621 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4681 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4741 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4801 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4861 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4921 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4981 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5041 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5101 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5161 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5221 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5281 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5341 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5401 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5461 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5521 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5581 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5641 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5701 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5761 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5821 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     5881 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5941 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6001 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6061 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6121 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6181 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6241 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6361 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6421 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6481 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6541 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6601 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6661 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6721 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6781 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6841 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6901 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6961 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7021 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7081 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7141 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7201 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7261 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7321 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7381 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7441 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7501 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7561 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7621 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7681 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7741 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7801 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7861 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7921 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7981 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8041 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8101 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8161 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8221 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8281 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8341 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     8401 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8461 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
