LOCUS       V021728                 6838 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021728
VERSION     V021728
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2358
                     /label="PAX8(NM_003466)"
                     /note="PAX8(NM_003466)"
                     /gene="PAX8"
     polyA_signal    2438..2662
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2708..3136
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3150..3479
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3546..4340
                     /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    4514..4635
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4684..4700)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4708..4724
                     /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(4732..4762)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4777..4798
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5086..5671)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5842..6702)
                     /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(6703..6807)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gcctcacaac
     1021 tccatcagat ctggccatgg agggctgaac cagctgggag gggcctttgt gaatggcaga
     1081 cctctgccgg aagtggtccg ccagcgcatc gtagacctgg cccaccaggg tgtaaggccc
     1141 tgcgacatct ctcgccagct ccgcgtcagc catggctgcg tcagcaagat ccttggcagg
     1201 tactacgaga ctggcagcat ccggcctgga gtgatagggg gctccaagcc caaggtggcc
     1261 acccccaagg tggtggagaa gattggggac tacaaacgcc agaaccctac catgtttgcc
     1321 tgggagatcc gagaccggct cctggctgag ggcgtctgtg acaatgacac tgtgcccagt
     1381 gtcagctcca ttaatagaat catccggacc aaagtgcagc aaccattcaa cctccctatg
     1441 gacagctgcg tggccaccaa gtccctgagt cccggacaca cgctgatccc cagctcagct
     1501 gtaactcccc cggagtcacc ccagtcggat tccctgggct ccacctactc catcaatggg
     1561 ctcctgggca tcgctcagcc tggcagcgac aagaggaaaa tggatgacag tgatcaggat
     1621 agctgccgac taagcattga ctcacagagc agcagcagcg gaccccgaaa gcaccttcgc
     1681 acggatgcct tcagccagca ccacctcgag ccgctcgagt gcccatttga gcggcagcac
     1741 tacccagagg cctatgcctc ccccagccac accaaaggcg agcagggcct ctacccgctg
     1801 cccttgctca acagcaccct ggacgacggg aaggccaccc tgaccccttc caacacgcca
     1861 ctggggcgca acctctcgac tcaccagacc taccccgtgg tggcagatcc tcactcaccc
     1921 ttcgccataa agcaggaaac ccccgaggtg tccagttcta gctccacccc ttcctcttta
     1981 tctagctccg cctttttgga tctgcagcaa gtcggctccg gggtcccgcc cttcaatgcc
     2041 tttccccatg ctgcctccgt gtacgggcag ttcacgggcc aggccctcct ctcagggcga
     2101 gagatggtgg ggcccacgct gcccggatac ccaccccaca tccccaccag cggacagggc
     2161 agctatgcct cctctgccat cgcaggcatg gtggcaggaa gtgaatactc tggcaatgcc
     2221 tatggccaca ccccctactc ctcctacagc gaggcctggc gcttccccaa ctccagcttg
     2281 ctgagttccc catattatta cagttccaca tcaaggccga gtgcaccgcc caccactgcc
     2341 acggcctttg accatctgta ggaattctgc agatatccag cacagtggcg gccgctcgag
     2401 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2461 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2521 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2581 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2641 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2701 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2761 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2821 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2881 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2941 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3001 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3061 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3121 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3481 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3541 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3601 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3661 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3721 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3781 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3841 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3901 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3961 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4021 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4081 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4141 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4201 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4261 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4321 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4381 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4441 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4501 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4621 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     5641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
