LOCUS       V043486                 8274 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043486
VERSION     V043486
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..2954
                     /label="SATB2(NM_001172509)"
                     /note="SATB2(NM_001172509)"
                     /gene="SATB2"
     CDS             2961..3026
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3050..3638
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3704..3752
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3954..4382
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4396..4725
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4792..5586
                     /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    5762..5883
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5932..5948)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5956..5972
                     /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(5980..6010)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6025..6046
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6334..6922)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7093..7953)
                     /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(7954..8058)
                     /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 ccaccatgga gcggcggagc gagagcccgt
      781 gtctgcggga cagccccgac cggcggagcg gcagcccgga cgtcaagggg cctcccccag
      841 tgaaggtggc ccggctggag cagaacggca gccccatggg agcccgcggg aggcccaacg
      901 gcgccgtggc caaggccgtg ggaggtttga tgattcctgt cttttgtgtc gtggagcagt
      961 tggacggctc tcttgaatat gacaacagag aagaacacgc cgagtttgtc ctggtgcgga
     1021 aagatgtgct ttttagccag ctggtggaga ctgcgctcct ggccctgggg tattctcaca
     1081 gctctgcggc ccaggcccaa ggaataatca agctgggaag gtggaaccct ctccccctca
     1141 gttatgtgac agatgcaccc gacgcgacag tggccgacat gctacaagat gtctatcatg
     1201 ttgtgacgtt gaaaatccaa ttacaaagtt gttcaaagtt ggaagacttg cctgcggagc
     1261 agtggaacca tgccacagtc cgcaatgcct taaaggaact gctcaaagag atgaaccaga
     1321 gcacattagc caaagaatgc cctctctccc agagtatgat ttcatccatt gtaaatagca
     1381 catattatgc caatgtgtca gcaaccaagt gccaggagtt tgggagatgg tataaaaagt
     1441 acaagaagat taaagtggaa agagtggaac gagaaaacct ttcagactat tgtgttctgg
     1501 gccagcgtcc aatgcattta ccaaatatga accagctggc atccctgggg aaaaccaacg
     1561 aacagtctcc tcacagccaa attcaccaca gtactccaat ccgaaaccaa gtgcccgcat
     1621 tacagcccat catgagccct ggtcttcttt ctccccagct tagtccacaa cttgtaaggc
     1681 aacaaatagc catggcccat ctgataaacc aacagattgc cgttagccgg ctcctggctc
     1741 accagcatcc tcaagccatc aaccagcagt tcctgaacca tccacccatc cccagagcag
     1801 ttaagccaga gccaaccaac tcttccgtgg aagtctctcc agatatctac cagcaagtca
     1861 gagatgagct gaagagggcc agtgtgtccc aagctgtctt tgcaagagtg gcattcaacc
     1921 gcacacaggg attgttgtct gagattctgc gtaaggaaga agaccctcgg acagcctctc
     1981 agtctcttct agtaaacctg agggccatgc agaatttcct caatctgcca gaagtggagc
     2041 gagatcgcat ctaccaggat gagagggagc ggagcatgaa tcccaatgtg agcatggtct
     2101 cctcggcctc cagcagtccc agctcctccc gaacccctca ggccaaaacc tcgacaccga
     2161 caacagacct ccctattaag gtggacggcg ccaacatcaa catcacagct gccatttatg
     2221 acgagatcca acaggagatg aaaagggcca aggtgtctca agccctgttt gccaaagtgg
     2281 ctgcaaataa aagtcagggc tggctgtgtg aactgctccg ctggaaggag aacccaagcc
     2341 cagaaaaccg caccctctgg gaaaacctct gtaccatccg tcgcttcctg aaccttcccc
     2401 agcatgagag ggatgtcatc tatgaggagg agtcaaggca tcaccacagc gaacgcatgc
     2461 aacacgtggt ccagcttccc cctgagccgg tgcaggtact tcatagacag cagtctcagc
     2521 cagccaagga gagttcccct cccagagaag aagcgcctcc cccacctcct ccgactgaag
     2581 acagttgtgc caaaaagccc cggtctcgca caaagatctc cttagaagcc ctggggatcc
     2641 tccaaagctt tattcatgat gtaggcctgt acccagacca ggaagccatc cacactcttt
     2701 cggctcagct ggatctcccc aaacacacca tcatcaagtt cttccagaac cagcggtacc
     2761 acgtgaagca ccacgggaag ctgaaagagc acctgggctc cgcggtggac gtggctgaat
     2821 ataaggacga ggagctgctg accgagtcag aggagaacga cagcgaggaa ggctccgagg
     2881 agatgtacaa agtggaggct gaggaggaaa atgctgacaa aagcaaggca gcacctgccg
     2941 aaattgacca gagaggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3001 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3061 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3121 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3181 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3241 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3301 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3361 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3421 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3481 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3541 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3601 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3661 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3721 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3781 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3841 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3901 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4321 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4561 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4621 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4681 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4741 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4801 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4861 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4921 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4981 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5041 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5101 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5161 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5221 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5281 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5341 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5401 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5461 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5521 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5581 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5641 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5701 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5761 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5881 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5941 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6001 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6061 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7261 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7621 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7921 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7981 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8041 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8101 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8161 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8221 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
