LOCUS       V029899                 7846 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029899
VERSION     V029899
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"
     CDS             964..3336
                     /label="Kank3(NM_030697)"
                     /note="Kank3(NM_030697)"
                     /gene="Kank3"
     CDS             3343..3408
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3446..3670
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3716..4144
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4158..4487
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4554..5348
                     /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    5522..5643
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5692..5708)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5716..5732
                     /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(5740..5770)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5785..5806
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6094..6679)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6850..7710)
                     /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(7711..7815)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcca agtttgtcct gaatcagaac cttcctgacc tgggcggccc tcccctgtac
     1021 ccaggcccca ctggaagtgc ccgcagccct agctctccct attctgtgga gacgccctat
     1081 ggcttccact tggacctgga cttcctcaaa tacgtggagg aaatcgagcg tggccctgcc
     1141 tctcgccgaa ctccaggacc tcctcatgca cgccgcccgc gcgcgtctcg aacgggcctc
     1201 gcgggagcgc ggagcccagg agcttggact tccagcgaat ccctggccag tgacgacggc
     1261 ggagcctcgg gtgcactctc tcctggtgcg ttcccggggc tctcgctacc tccgctgtcg
     1321 ccacgctcct tgtcgcgcaa tccgcgcgtt gagcacacgc tcttggagac cagccgacga
     1381 ctggagcagg cgcaggctcg ggagcgcgcg ctcagcccgg cccgcgcagt cacacgcagt
     1441 ccgcgcgggt ccggccgtag cagccccgcc cctaaccccg cccttgcctc cccgggccct
     1501 gcccagctgc agctagtgag ggagcagatg gccgcggcgc tgcggcgcct gcgtgagctc
     1561 gaggaccagg cgcgcgctct gcccgagctg caggagcagg tgcgcgcgct gcgtgccgag
     1621 aaggcgcggc tactggccgg gcgcgtgcag ccagaacaag aagtagagat cgaggcgcgc
     1681 ccagacaagc tcgcccagct tcggcgcctc accgagcgtc tggccacctc ggatcgtgga
     1741 gtgcgctcca gggctagccc ccgggcagag gatcccgacg ggttggctgc caggcgcagc
     1801 gaaggagcgc tgcaggtgct cgacccgggg tctaggacac ccgatgggga accccggact
     1861 cgggagacgg gcaccgaagt cgtgcctgag acccgagagg tggatgccca ggcagtaccc
     1921 gagacagggg aggctggagt ggaggtggtc cctgagaccg ttgaggtgga cacgtgggtg
     1981 actgaggagc tcctagggtt gcctgaggct gcagagcgtg agctggagct gctccgcacc
     2041 agcttggagc atcagcgagg ggtgagcgaa ctcttgcggg gccgattgag ggagttggag
     2101 gaggcccacg aggccgcagt gaccaggcct cagtctcgcg acgttgctgc ccagactaca
     2161 ttgggttgca ctgagaaaac cacacagaca gagctgccag tggagaacca gcccagaccc
     2221 acggctggag acgagatggc ccctgttggc atcctcaaat ccatcatgaa gaagaaagat
     2281 ggtattccag gtgcacaatc cagtcaggga cccaagagtc tgcagttcgt tggggtcctc
     2341 aacggagagt atgagagttc ctccagtgag gatggcaaca gcgatgatga agatggtgtt
     2401 gctgaacacc ccaggagtag ctcttctgga tcagatgata gcagtggggg atctgacgct
     2461 ggaacccctg gcccccacaa tgacaaagat gctggggact gcgagcttga gacacatcca
     2521 gagctgacgg cagggagaga agggaggtgt gaactgaacc cccgtttgag ggaggcttgc
     2581 attgctctga atcagcagct gaaccggcca cgtggagtca ccagcagaga tggcaatgca
     2641 gcacgccttg tggcccagga atggtttcga gtgtccagcc aaaaacgctc tcaggcagag
     2701 tctgtggctg gggttcttcg aggggtgaaa agcctggggc ctgagctgct ggcttatgtg
     2761 gtgaacctgg ctgatggcaa tggaaacaca gctctgcatt acagcgtgtc tcatgggaat
     2821 ctcgccattt ccagcctgct actggataca ggggtctgtg atgtgaatca ccagaataga
     2881 gctggctact ctgccctcat gttggctgca ctcacttctg taggacaaga ggaggaagac
     2941 atggctgtgg cccagagact tttcagtatg ggtgatgtca atgccaaggc tagccagact
     3001 ggacagacag ccctcatgct ggccatcagc catggccacc aggacatggt agctgccttg
     3061 ctagaatgcg gggctgatgt caatgtacag gatgctgatg gggccacagc actgatgtgt
     3121 gccagcgagt atggacgcct tgatacagtt caattgttgt tggcccagcc aggttgtgac
     3181 ctaaccatct tagacaatga gggcacaagt gccctggcca ttgcactgga ggctgaacag
     3241 gatgaggtgg ctgcactgct ccatgcccac ctgacatcaa accaccaggg acagtcttcc
     3301 acagggtctc ccactgcaaa ggaatgcaat gacaaactcg aggactacaa agaccatgac
     3361 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     3421 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3481 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3541 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3601 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3661 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3721 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3781 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3841 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3901 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3961 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4021 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4081 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4141 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4321 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4381 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4441 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4501 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4561 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4621 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4681 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4741 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4801 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4861 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4921 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4981 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5041 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5101 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5161 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5221 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5281 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5341 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5401 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5461 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5521 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5581 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5641 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5821 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5881 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6601 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
