LOCUS       V043741                 6919 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043741
VERSION     V043741
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..1923
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID
                     engineered for improved catalytic efficiency (Branon et
                     al., 2018)"
                     /transl_table=1
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
                     /label="TurboID"
     CDS             1954..2409
                     /label="NINJ1(NM_004148)"
                     /note="NINJ1(NM_004148)"
                     /gene="NINJ1"
     CDS             2416..2481
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2519..2743
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2789..3217
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3231..3560
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3627..4421
                     /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    4595..4716
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4765..4781)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4789..4805
                     /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(4813..4843)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4858..4879
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5167..5752)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5923..6783)
                     /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(6784..6888)
                     /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 accatgaaag acaatactgt gcctctgaag ctgatcgctc tcctggctaa tggcgagttc
     1021 catagtggcg aacagctggg agaaaccctg ggcatgtcca gggccgctat caacaagcac
     1081 attcagactc tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa gggctactct
     1141 ctgcccgagc ctatcccgct gctgaacgct aaacagattc tgggacagct ggacggcggg
     1201 agcgtggcag tcctgcctgt ggtcgactcc accaatcagt acctgctgga tcgaatcggc
     1261 gagctgaaga gtggggatgc ttgcattgca gaatatcagc aggcagggag aggaagcaga
     1321 gggaggaaat ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg
     1381 aagcggggac cagcagcaat cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg
     1441 ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag
     1501 gatagaaagc tggcaggcat cctggtggag ctggccggaa taacaggcga tgctgcacag
     1561 atcgtcattg gcgccgggat taacgtggct atgaggcgcg tggaggaaag cgtggtcaat
     1621 cagggctgga tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct
     1681 acgctgatcc gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca
     1741 tatctgccac ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt
     1801 ggggacaaag agattttcgg gattagccgg gggattgata aacagggagc cctgctgctg
     1861 gaacaggacg gagttatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa
     1921 aagggtggag gcggatcagg aggcggtgga tctatggact cgggaaccga ggagtacgag
     1981 ctcaacggcg gcctgcctcc gggcacaccc ggctccccgg acgcctcgcc ggcccgctgg
     2041 ggctggaggc acgggcccat caacgtgaac cattacgcca gcaagaagag cgcagccgag
     2101 agcatgctgg acatcgcgct gctgatggcc aacgcgtccc agctgaaggc cgtcgtggaa
     2161 cagggcccca gcttcgcctt ctatgtgccc ctggtggtcc tcatctccat ctcccttgtg
     2221 ctgcagatcg gcgtgggggt gctgctcatc ttccttgtca agtacgacct taacaacccg
     2281 gccaagcacg ccaagctgga cttcctcaac aacctggcca cgggcctggt gttcatcatc
     2341 gtggtagtca acatcttcat cacggccttc ggggtccaga agcccttgat ggacatggca
     2401 ccccagcagc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     2461 tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc agcctcgact
     2521 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2581 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2641 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2701 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2761 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2941 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3001 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3061 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3121 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3181 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3241 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3301 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3361 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3421 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3481 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3541 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3601 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3661 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3721 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3781 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3841 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3901 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3961 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4021 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4081 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4141 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4201 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4261 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4321 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4381 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4441 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4501 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4561 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4621 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4681 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4741 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5581 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5701 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5941 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6001 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6061 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6121 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6181 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6241 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6301 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6361 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6421 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6481 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6541 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6601 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6661 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6721 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6781 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6841 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6901 aaaagtgcca cctgacgtc
//
