LOCUS       V022435                 7925 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022435
VERSION     V022435
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..2605
                     /label="HSPA1A(NM_005345)"
                     /note="HSPA1A(NM_005345)"
                     /gene="HSPA1A"
     CDS             2612..2677
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2701..3289
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3355..3403
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3605..4033
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4047..4376
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4443..5237
                     /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    5413..5534
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5583..5599)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5607..5623
                     /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(5631..5661)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5676..5697
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5985..6573)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6744..7604)
                     /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(7605..7709)
                     /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 ccatggccaa agccgcggcg atcggcatcg acctgggcac
      721 cacctactcc tgcgtggggg tgttccaaca cggcaaggtg gagatcatcg ccaacgacca
      781 gggcaaccgc accaccccca gctacgtggc cttcacggac accgagcggc tcatcgggga
      841 tgcggccaag aaccaggtgg cgctgaaccc gcagaacacc gtgtttgacg cgaagcggct
      901 gattggccgc aagttcggcg acccggtggt gcagtcggac atgaagcact ggcctttcca
      961 ggtgatcaac gacggagaca agcccaaggt gcaggtgagc tacaaggggg agaccaaggc
     1021 attctacccc gaggagatct cgtccatggt gctgaccaag atgaaggaga tcgccgaggc
     1081 gtacctgggc tacccggtga ccaacgcggt gatcaccgtg ccggcctact tcaacgactc
     1141 gcagcgccag gccaccaagg atgcgggtgt gatcgcgggg ctcaacgtgc tgcggatcat
     1201 caacgagccc acggccgccg ccatcgccta cggcctggac agaacgggca agggggagcg
     1261 caacgtgctc atctttgacc tgggcggggg caccttcgac gtgtccatcc tgacgatcga
     1321 cgacggcatc ttcgaggtga aggccacggc cggggacacc cacctgggtg gggaggactt
     1381 tgacaacagg ctggtgaacc acttcgtgga ggagttcaag agaaaacaca agaaggacat
     1441 cagccagaac aagcgagccg tgaggcggct gcgcaccgcc tgcgagaggg ccaagaggac
     1501 cctgtcgtcc agcacccagg ccagcctgga gatcgactcc ctgtttgagg gcatcgactt
     1561 ctacacgtcc atcaccaggg cgaggttcga ggagctgtgc tccgacctgt tccgaagcac
     1621 cctggagccc gtggagaagg ctctgcgcga cgccaagctg gacaaggccc agattcacga
     1681 cctggtcctg gtcgggggct ccacccgcat ccccaaggtg cagaagctgc tgcaggactt
     1741 cttcaacggg cgcgacctga acaagagcat caaccccgac gaggctgtgg cctacggggc
     1801 ggcggtgcag gcggccatcc tgatggggga caagtccgag aacgtgcagg acctgctgct
     1861 gctggacgtg gctcccctgt cgctggggct ggagacggcc ggaggcgtga tgactgccct
     1921 gatcaagcgc aactccacca tccccaccaa gcagacgcag atcttcacca cctactccga
     1981 caaccaaccc ggggtgctga tccaggtgta cgagggcgag agggccatga cgaaagacaa
     2041 caatctgttg gggcgcttcg agctgagcgg catccctccg gcccccaggg gcgtgcccca
     2101 gatcgaggtg accttcgaca tcgatgccaa cggcatcctg aacgtcacgg ccacggacaa
     2161 gagcaccggc aaggccaaca agatcaccat caccaacgac aagggccgcc tgagcaagga
     2221 ggagatcgag cgcatggtgc aggaggcgga gaagtacaaa gcggaggacg aggtgcagcg
     2281 cgagagggtg tcagccaaga acgccctgga gtcctacgcc ttcaacatga agagcgccgt
     2341 ggaggatgag gggctcaagg gcaagatcag cgaggcggac aagaagaagg tgctggacaa
     2401 gtgtcaagag gtcatctcgt ggctggacgc caacaccttg gccgagaagg acgagtttga
     2461 gcacaagagg aaggagctgg agcaggtgtg taaccccatc atcagcggac tgtaccaggg
     2521 tgccggtggt cccgggcctg ggggcttcgg ggctcagggt cccaagggag ggtctgggtc
     2581 aggccccacc attgaggagg tagatctcga ggactacaaa gaccatgacg gtgattataa
     2641 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     2701 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2761 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2821 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2881 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2941 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3001 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3061 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3121 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3181 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3241 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     3301 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3361 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3421 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3481 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3541 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     3601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3961 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4381 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4441 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4621 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4981 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5221 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5281 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5341 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5401 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5521 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5581 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5641 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5701 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5761 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5821 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5881 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5941 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6001 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6061 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6121 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6181 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6241 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6301 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6361 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6421 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6481 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6541 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6601 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6661 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6721 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6781 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6841 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6901 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6961 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7021 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7081 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7141 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7201 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7261 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7321 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7381 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7441 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7501 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7561 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7621 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7681 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7741 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7801 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7861 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7921 ttagg
//
