LOCUS       V021838                 8825 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021838
VERSION     V021838
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..3472
                     /label="Pcdhga4(NM_033587)"
                     /note="Pcdhga4(NM_033587)"
                     /gene="Pcdhga4"
     CDS             3512..3577
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3601..4189
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4255..4303
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4505..4933
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4947..5276
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5343..6137
                     /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    6313..6434
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6483..6499)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6507..6523
                     /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(6531..6561)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6576..6597
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6885..7473)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7644..8504)
                     /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(8505..8609)
                     /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 ccatggcggc tccttacaaa tcagaccgca ggggactaat
      721 ctggatctgc attttccttg gatccctatg cgacatccgc gcggaacaga tccggtactc
      781 tgtgccggag gagctggaaa ggggctctgt ggtgggtaat ctcgccgcgg acctggggct
      841 ggagcctgga aaactggcag aacgcggagt ccgcatcgtc tccagaggta agacccagct
      901 ctttgctctg aacccgcgaa gcggcagttt ggtgacagca gggagggtcg acagagaagg
      961 gctctgtgac agatccccaa agtgtaccgc aaatctggag attctcctag aggataaagt
     1021 gaggattcta gccatagaag tggagattat cgatgttaat gataatgccc ccagctttgg
     1081 agcacagcag agagaaataa aagtcgctga aagtgaaaat cctgggacaa gatttcctct
     1141 tcctgaagcc tttgatctgg acataggggt gaacgccttg cagggttacc agctcagctc
     1201 caatgatcac ttctccctgg atgtgcaaag tggaccggat gggattaagt atccagagct
     1261 agtgctggaa aatgccttag accgcgagga agaagccgtt catcacctgg ttctcaccgc
     1321 cttcgacgga ggcgacccgg ttcgctctgg tactgccaca attcaagtaa cactcgtgga
     1381 caccaacgac aatgctcctg tattcactca gccagagtac cacattagtg tgaaagagaa
     1441 tttgcctgta ggcacccggc tgcttaccat aaaagccact gacccggatg aaggagtcaa
     1501 cggagaagtg acgtattctt tccggaacgt aagagagaaa atctcccagc tcttccagct
     1561 gaattctctg actggggaca ttacagtgtt gggggaactg gattatgaag actctggatt
     1621 ctacgatgtt gacgtagaag cccatgatgg gcctggtctt cgagccagaa gtaaggtact
     1681 ggtgacagtt ctggatgtaa atgacaatgc cccggaagtc acagtcacct ctctcaccag
     1741 ctctatccag gaagcttctt ccccaggcac agtgattgca ctcttcaacg tgcacgacag
     1801 cgactctgga gagaacggcc ttgtcacttg ttctattcca gataatcttc cattcagact
     1861 ggaaaagacc tatggaaatt atcatcggtt gttgatacac aggactctgg acagggaaga
     1921 agtctctgac tataacatta caataactgc caccgaccag ggaactccac ctctgtctac
     1981 agagacttat atctcattgc aggtggtaga tattaatgac aacccgccga ccttcaccca
     2041 tgcctcctat tctgcttaca ttccagaaaa caaccctaga ggagcttcca tcttatccat
     2101 aacagcccag gacccagaca gtggtgagaa tgcccaggtt atctattctc tgtcagagga
     2161 cactatccag ggggcaccta tgtcctccta cgtctccata aactccaata ccggtgtcct
     2221 ttatgcactg cgttcctttg attatgagca gtttcaagac ttgaagttac tggtgacagc
     2281 cagggacagt gggactcctc cactcagcag caacgtgtcc ctgagcttgt ctgtgctgga
     2341 ccagaacgat aacacccctg agatcctgta tcccaccatc cccactgatg gttctactgg
     2401 tgtggagctg acacctcgtt ctgcagatcc tggatacctg gtgaccaagg tggtggccgt
     2461 agacaaagac tcaggacaga acgcctggct gtcctaccgc ctgctcaagg ccagcgagcc
     2521 agggctcttc tcagtggggc tgcacacagg tgaggtacgc actgctcggg ccctgctgga
     2581 cagagatgct ctcaagcaga gcctggtggt gaccgtgcag gaccatggcc agccccctct
     2641 ctccgccact gtcacactca ccatagctgt gtcagacaac attccagaca tcctggctga
     2701 tctggtcaac attaacgccc ccattgacca ggaggattca gacatcacat tgtacctggt
     2761 ggtagctgtg gctgcggtct cttgtgtctt cttagccttt gtcattgtgc tgctgataca
     2821 caggctccgt cgctggcact caactcgcct gcttcaggct gcaggtaatg ggttgtcatc
     2881 cttgcctgcc tcacattttg tgggtgtgga tggtgtacac gctttcctac agacctattc
     2941 tcatgaggtc tcactcacag cagactctgg gaagagtcac ctgatcttcc cacagcccaa
     3001 ctacgcagac acactcatca gccaggagag ctgtgggaag agtgatcctc tcctggtatc
     3061 tcaagacttg cttgaaataa aaggagactc cagtctgcag caagccccgc ccaacactga
     3121 ctggcgtttc tctcaagccc agagacccgg cacgagcgga tcccaaaatg gtgatgaaac
     3181 tggcacctgg cccaacaacc agtttgatac agagatgctg caagccatga tcttggcctc
     3241 tgccagcgaa gctgctgatg ggagctctac cctgggaggg ggcgctggca ccatgggtct
     3301 gagcgctcga tacggacccc agtttaccct gcagcacgtg cctgactacc gccagaacgt
     3361 gtacatccct ggcagcaatg ccacgctgac caatgccgct ggcaaacgag atggcaaggc
     3421 tccagcaggt ggtaatggca acaagaagaa atcgggcaag aaagagaaga aggaattctg
     3481 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa
     3541 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     3601 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3661 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3721 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3781 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3841 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3901 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3961 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4021 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4081 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4141 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     4201 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4261 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4321 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4381 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4441 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4501 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4561 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4801 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4861 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4921 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4981 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5041 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5101 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5161 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5221 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5281 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5341 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5401 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5461 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5521 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5581 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5641 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5701 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5761 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5821 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5881 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5941 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6001 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6061 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6121 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     6181 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6241 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6301 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6361 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6421 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6481 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     7081 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7321 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8641 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     8701 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     8761 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8821 ttagg
//
