LOCUS       V043876                 6707 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043876
VERSION     V043876
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"
     protein_bind    820..838
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    841..859
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     CDS             1008..2585
                     /label="IKZF2(NM_016260)"
                     /note="IKZF2(NM_016260)"
                     /gene="IKZF2"
     polyA_signal    2665..2889
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    3173..3220
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2μ plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /label="FRT"
     CDS             3228..4250
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     *"
                     /label="HygR"
     polyA_signal    4380..4501
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4550..4566)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4574..4590
                     /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(4598..4628)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4643..4664
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4952..5540)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5711..6571)
                     /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(6572..6676)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cgccaccatg gaaacagagg
     1021 ctattgatgg ctatataacg tgtgacaatg agctttcacc cgaaagggag cactccaata
     1081 tggcaattga cctcacctca agcacaccca atggacagca tgcctcacca agtcacatga
     1141 caagcacaaa ttcagtaaag ctagaaatgc agagtgatga agagtgtgac aggaaacccc
     1201 tgagccgtga agatgagatc aggggccatg atgagggtag cagcctagaa gaacccctaa
     1261 ttgagagcag cgaggtggct gacaacagga aagtccagga gcttcaaggc gagggaggaa
     1321 tccggcttcc gaatggtaaa ctgaaatgtg acgtctgtgg catggtttgc attgggccca
     1381 atgtgcttat ggtacataaa aggagtcaca ctggtgaacg ccccttccac tgtaaccagt
     1441 gtggagcttc ttttactcag aagggcaacc ttctgagaca cataaagtta cactctggag
     1501 agaagccgtt caaatgtcct ttctgtagct acgcctgtag aagaagggac gccctcacag
     1561 gacacctcag gacccattct gtgggtaaac ctcacaagtg caactactgt ggacgaagct
     1621 acaagcagcg cagttcactg gaggagcaca aggaacgctg ccacaactat ctccagaatg
     1681 tcagcatgga ggctgctggg caggtcatga gtcaccatgt acctcctatg gaagattgta
     1741 aggaacaaga gcctattatg gacaacaata tttctctggt gccttttgag agacctgctg
     1801 tcatagagaa gctcacgggg aatatgggaa aacgtaaaag ctccactcca caaaagtttg
     1861 tgggggaaaa gctcatgcga ttcagctacc cagatattca ctttgatatg aacttaacat
     1921 atgagaagga ggctgagctg atgcagtctc atatgatgga ccaagccatc aacaatgcaa
     1981 tcacctacct tggagctgag gcccttcacc ctctgatgca gcacccgcca agcacaatcg
     2041 ctgaagtggc cccagttata agctcagctt attctcaggt ctatcatcca aataggatag
     2101 aaagacccat tagcagggaa actgctgata gtcatgaaaa caacatggat ggccccatct
     2161 ctctcatcag accaaagagt cgaccccagg aaagagaggc ctctcccagc aatagctgcc
     2221 tggattccac tgactcagaa agcagccatg atgaccacca gtcctaccaa ggacaccctg
     2281 ccttaaatcc caagaggaaa caaagcccag cttacatgaa ggaggatgtc aaagctttgg
     2341 atactaccaa ggctcctaag ggctctctga aggacatcta caaggtcttc aatggagaag
     2401 gagaacagat tagggccttc aagtgtgagc actgccgagt ccttttccta gaccatgtca
     2461 tgtacaccat tcacatgggt tgccatggct accgggaccc actggaatgc aacatctgtg
     2521 gctacagaag ccaggaccgt tatgagtttt catcacacat tgttcgaggg gagcacacat
     2581 tccacgtgag cggctggcgg ctgttcaaga agattagcta gctcgagtct agagggcccg
     2641 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2701 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2761 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2821 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2881 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2941 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3001 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3061 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3121 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     3181 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     3241 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     3301 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     3361 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     3421 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     3481 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     3541 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     3601 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     3661 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     3721 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     3781 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     3841 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     3901 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     3961 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4021 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4081 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     4141 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     4201 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     4261 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4321 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4441 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4501 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
