LOCUS       V018870                 7231 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018870
VERSION     V018870
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             997..2784
                     /label="Zc3h12a(NM_153159)"
                     /note="Zc3h12a(NM_153159)"
                     /gene="Zc3h12a"
     polyA_signal    2831..3055
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3101..3529
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3543..3872
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3939..4733
                     /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    4907..5028
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5077..5093)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5101..5117
                     /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(5125..5155)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5170..5191
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5479..6064)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6235..7095)
                     /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(7096..7200)
                     /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 ggtaccatga gtgacccttg tggaacgaag
     1021 cctgtccaag aatccaaccc caccatgagt ctgtggagtc ttgaggacag acacagcagc
     1081 cagggtcgac ctcagccaga ccaggatcct gtggctaaag aggcccctac ttccgagctg
     1141 cagatgaagg tggacttttt ccgtaaactg gggtactcgt cctctgagat ccacagtgtc
     1201 ctgcagaagc tgggagtcca agcagacacc aacacggtgc taggggaatt ggtgaagcat
     1261 ggctcagcta ctgaacgaga atgccaggcc ctgacggccc ccagccccca gccccctctg
     1321 gtgccccggg gtggaagcac ccccaagcct tccactctag aaccctcact cccagaggag
     1381 gacagagagg gcagcgacct gagacctgtg gtcatcgacg gaagcaatgt ggccatgagc
     1441 catgggaaca aggaagtctt ctcttgccgg ggcattctgc tggctgtgaa ctggtttctg
     1501 gagcgaggcc acacagatat taccgtgttt gtgccatctt ggaggaagga acagcctcga
     1561 ccagatgtgc ctatcacaga ccagcacatc cttcgggaac tagagaaaaa gaagatcttg
     1621 gtgttcacgc catccaggcg ggtcggcggc aagcgcgtgg tgtgctatga tgaccgcttc
     1681 attgtgaagc tggccttcga atccgacgga gtggtggtct ccaatgacac gtaccgagac
     1741 ctccaaggcg agaggcagga gtggaaacgc ttcatcgagg agcggctgct catgtactcc
     1801 ttcgtcaatg acaagttcat gccccctgac gaccctttag gacggcatgg gcctagcctg
     1861 gacaacttcc ttcgtaagaa accactgcct tctgagcaca ggaagcagcc atgcccctat
     1921 gggaagaaat gtacgtatgg aatcaagtgc cgatttttcc accctgagcg gccaagccgt
     1981 ccccagcgct ctgtggccga tgagctccgt gccaacgctc tcctctcacc tcccaggact
     2041 ccagtcaagg acaaaagtag ccagaggcct tcccctgcct ctcagtccag ctctgtgtcc
     2101 ctagaggctg aaccaggcag cctggatggg aaaaagctgg gtgccaggtc atctccgggt
     2161 ccccaccgag aaggctcacc gcagacctgt gctccagctg gcaggagcct ccctgttagt
     2221 gggggcagct ttgggcccac agagtggctt gcacacaccc aggactcact cccatacacc
     2281 tcccaggagt gccttgattc aggcattggt tccctggaga gccagatgtc agaattatgg
     2341 ggcgtgcgag gaggcagccc tggggagtcg ggccccactc ggggccccta tgcaggttat
     2401 cacagctatg gatccaaggt cccagcagca ccttcctttt ctccttttag accagccatg
     2461 ggtgctggcc acttcagtgt ccccaccgac tatgtgcccc cgccacccac ctacccatcc
     2521 agagagtact ggtctgagcc gtacccatta cccccaccca ctcctgtcct tcaggagccc
     2581 cagagaccca gccccggggc tggtgggggc ccctggggca gggtgggtga cctggccaaa
     2641 gaaagggctg gtgtatatac caagctgtgt ggtgtcttcc ccccacacct ggtagaagct
     2701 gtaatgagac gcttcccaca gctgctggat ccgcagcagc tggccgcaga gatcctgtct
     2761 tacaagtccc agcacctcag tgagtaactc gagtctagag ggcccgttta aacccgctga
     2821 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2881 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2941 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3001 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3061 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3121 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3181 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3241 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3301 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3361 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3541 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3601 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3661 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3721 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3781 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3841 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3901 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3961 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4021 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4081 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4141 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4201 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4261 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4321 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4381 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4441 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4501 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4561 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4621 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4681 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4741 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5041 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5101 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5161 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5221 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5281 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6001 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7201 cacatttccc cgaaaagtgc cacctgacgt c
//
