LOCUS       V033550                 9066 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033550
VERSION     V033550
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3746
                     /label="Snx19(NM_028874)"
                     /note="Snx19(NM_028874)"
                     /gene="Snx19"
     CDS             3753..3818
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3842..4430
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4496..4544
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4746..5174
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5188..5517
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5584..6378
                     /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    6554..6675
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6724..6740)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6748..6764
                     /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(6772..6802)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6817..6838
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7126..7714)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7885..8745)
                     /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(8746..8850)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgaa ggcacaaaca gtgtccccta
      781 cccaggggac tatctccgag tccagttatg tccacagcaa tctatggagt agccggaagc
      841 taatgatcgt gggggtcttg gtgggctggc tactggtcat ccaccttctg gtcaacatgt
      901 ggcttctgat ccttctatgt gcatcgctgg tagcgctggg aggatggctg ggctccactg
      961 ccatcctagg ggcttcaggt caattgcacc tggaacgatt cattaccatc accacctgtc
     1021 ctccatgtcc cgaggcagaa aggcagctgg aacaggagat taaccgcact atccagatga
     1081 ttatccgaga cttcgtgtta tcctggtatc gttctgtgag ccatgagccg gccttcgagg
     1141 cagagatgga ggcagccatg aaagggttgg ttcaggagct tcgaaggcgg atgagcatag
     1201 tggacagcca tgctcttacc cagagggttc tgactctctg tggttgtcac ctgcagagct
     1261 atattcaggc aaaggaggct actgcaaagg agcagagctg tccagttcag ccctcccagc
     1321 tgtgggatgc ttactgccag gttacagctc cccatcctgc catgagctgt cccaccacgg
     1381 aggtcactta tgcacgtggc attgtgaact tgatactaaa agagttagtg cccaagcccc
     1441 acctggagac gaggactgga cgccacgttg tggttgaagt tattacttgc aatgtcatct
     1501 taccactgat cagcaagcta tcagatcctg actggatcca ccttattctt gtgagtatct
     1561 tttccaagta cagacatgat gcagcacagg gaactaaacc cccctgctca tccagtgtcc
     1621 tagagcagcc ctcggtgccc acatctctgc cattgattgt tgaagtagag agtctgccag
     1681 taggaaaagc atcttctcca gcaacagccc cagtacactt gacctccagt gagccagccc
     1741 cttccccaga gattgaagaa ggccatgaag ctgtagaggg agatttgcct gggatgctcg
     1801 aagaaaaaaa agtaggaaat agctcatccc attttctaca gccagatatc cgaggcccct
     1861 tgttcttatg tgaagactca gaactggagt caccactgtc tgaactgagc aaagaaacca
     1921 tcctgctaat gaccccaggc aacttccttt ctgacaggat tcaagatgct ctgtgtgccc
     1981 tagacgactc cggggctctg gagcctaagg atggcgaggg atctgaatgt atggaaggag
     2041 cagaggccga ggaggcccca gggacagaca cagagacagg catgctggtc tccgtgctta
     2101 attgcccaga gatccagatt gacacagcag acaaggaggt ggagcaagga gatgacacct
     2161 ctcttacagc tttgctggag gaaccagaaa aaccctgccc cctacggccc tcatgtttag
     2221 acaaagatct cgccagtggt gtgtgctccc ttgaacctgc tatgccccca gtgcctcttt
     2281 cctcctctcc acctggtcct ctcagctcag ccaccttcag cttcgagtcc ctgagcagtc
     2341 cagacggccc ggttgtcatc cagaaccttc gaatcactgg caccatcact gcccgagagc
     2401 acagtgggac tggattccac ccatacacac tgtacactgt gaagtacgag actgtcctta
     2461 atggcgagaa cagcagcggc ctacagcagc tggcctatca caccgtgaac cgccgctacc
     2521 gggagttctt gaatctgcag acccgcctgg aggagaaacc agatctacga aagtttatca
     2581 aaaatgtgaa gggtccaaaa aagctctttc cagatcttcc gtttggaaac atggatagtg
     2641 acagagtaga agccaggaag agcctcctgg agtcattcct gaagcaactc tgcgccattc
     2701 ctgagattgg caacagtgag gaggtgcagg agttccttgc tctgaacaca gatgctcgga
     2761 ttgcctttgt caagaaacca ttcatggtct ctaggataga caagatggtg gtgagtgcta
     2821 tcgtggacac cttgaagaca gcatttcctc gctctgaacc ccagagtccc acagaggagc
     2881 tgagtgaagc agagaatgaa agcaagcccc agacagaagg caagaaggct agcaagtcca
     2941 gactgaggtt ttcatccagt aaaattgccc ctgcactgag tatagccgag gcacaggaca
     3001 agattctgta ctgcctccag gaaggcaatt cggagtcaga agtcctgtcc atgtctggga
     3061 tggagtcttt cattgaaaaa cagacaaagt tactgagaat ccagccagca gaagtcccgg
     3121 ataaagatcc ccagcaagtg cccaaagaat atgtggatag cggtctgctg gacaaagctg
     3181 tggtagccca agagctcaac aagagtggcc cgggaacgga gacagagtta gccgacacgg
     3241 cctttgatct gatcctcctg ctgttgatgg agcaatggaa gtggctgtgt actgaaagca
     3301 tgcagaagtt cctccacatt atttttggca cactagttca gaggtggcta gaagtgcagg
     3361 tggctaatct aacatgtccc cagcgctggg cacagtacct ccatcttctt cgggaatcta
     3421 tctggcctgg tggagtgctg cccaagtttc cacggcctgg aaggacgcag gcacagaaag
     3481 cagctacgga gaagcaggct ttgcagagct tgatggatct ccttccagat tttttggtag
     3541 aaatcctggg agtgaacaag tgtcggctga gctggagtct tgtcttggag tcattccagc
     3601 agcccctcat caacagacat ttgatttact gccttggaga cattatcctg gagttgctgg
     3661 acctcagtgc ctctgttgaa gagtgtgccc cagctacctc tgcctcagat tccccaggaa
     3721 gcctcaagaa gatggctgtc tccactggat ccgactacaa agaccatgac ggtgattata
     3781 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3841 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3901 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3961 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4021 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4081 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4141 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4201 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4261 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4321 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4381 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     4441 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4501 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4561 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4621 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4681 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4741 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4801 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4861 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4921 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4981 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5041 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5101 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5161 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5221 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5281 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5341 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5401 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5461 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5521 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5581 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5641 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5701 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5761 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5821 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5881 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5941 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6001 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6061 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6121 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6181 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6241 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6301 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6361 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     6421 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6481 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6541 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6601 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6661 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6721 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6781 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7321 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7561 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8881 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8941 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9001 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9061 cttagg
//
