LOCUS       V047411                 8392 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047411
VERSION     V047411
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..3882
                     /label="PJA1(NM_145119)"
                     /note="PJA1(NM_145119)"
                     /gene="PJA1"
     CDS             3889..3954
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3992..4216
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4262..4690
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4704..5033
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5100..5894
                     /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    6068..6189
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6238..6254)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6262..6278
                     /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(6286..6316)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6331..6352
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6640..7225)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7396..8256)
                     /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(8257..8361)
                     /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 tctatgggtc aggaatctag caagcctgta
     1981 tggcccaatc caacaggagg gtatcagtcc aatacaggta ggaggtatgg aagaaggcat
     2041 gcttatgtca gtttcaggcc acccacgagc cagcgggaaa ggattgccag ccagagaaag
     2101 acgaactccg aagtcccaat gcacagatca gcccccagtc aaaccaccaa gaggagccga
     2161 tcgccatttt ccactactcg tcgtagttgg gacgacagcg agagttcggg aaccaacctg
     2221 aatattgata atgaggacta ttccaggtat ccgccaagag agtacagagc ttcgggtagc
     2281 agaagaggaa tggcctatgg acatattgac tcttatgggg cagatgatag tgaggaggag
     2341 ggggctgggc ctgttgagcg accgccagtg agagggaaaa ctggcaagtt taaagatgat
     2401 aagctgtatg acccagagaa aggggcaagg tctttggctg ggccacctcc acatttctct
     2461 agttttagcc gtgatgtgag agaggagcga gacaagttag acccagtccc tgcagcaaga
     2521 tgctcagcta gcagagctga cttcctgcca caaagtagtg tggcctcaca gtcgtcttct
     2581 gaaggcaagc tggctacaaa aggtgacagc tcggagaggg agagaaggga gcaaaattta
     2641 cctgcacgtc ccagcagggc tcctgtgagt atttgtggtg gtggggaaaa cacctcaaag
     2701 agtgcagagg aacctgtggt caggcccaaa atcagaaacc tggcaagtcc aaactgcgtg
     2761 aaaccaaaaa ttttttttga tactgatgat gatgacgata tgccacacag tacttccagg
     2821 tggagggata ccgccaatga caatgagggc cactcggatg gcctggcaag aagagggaga
     2881 ggcgagagtt caagtggcta tcccgagcca aagtaccctg aagacaaacg ggaagcgagg
     2941 agtgaccaag tgaaaccaga aaaggtgccg agacgacgac gcaccatggc cgaccctgac
     3001 ttctggacgc acagtgatga ttactacaaa tactgcgacg aagactctga cagtgacaaa
     3061 gagtggattg ctgctctgcg tcggaaatat cgaagccgag agcaaaccct gtcctccagt
     3121 ggcgaaagct gggagactct gccggggaaa gaagagcggg aacctccaca ggctaaggtg
     3181 agtgccagca ctggcaccag ccctggcccc ggtgctagtg ccagtgccgg ggctggcgcc
     3241 ggggccagtg ctggcagcaa tggcagcaat taccttgaag aagttcgaga accatctctt
     3301 caggaagagc aggcatccct ggaagaagga gaaattcctt ggctccagta ccatgagaat
     3361 gacagtagca gtgaggggga taatgattct ggtcacgagt tgatgcaacc tggggtattc
     3421 atgctggatg gaaacaacaa ccttgaagat gactccagtg tgagcgaaga cctagaagtg
     3481 gattggagcc tctttgatgg atttgcagat gggttaggag tggctgaagc catttcctat
     3541 gtggaccctc agttcctcac ctacatggca cttgaagaac gcctggccca ggcaatggaa
     3601 actgcccttg cgcacttgga gtctctcgca gtggatgtag aggtggccaa tccaccagca
     3661 agcaaggaga gcattgacgc tcttcccgag atcctggtca ctgaagatca tggcgcagtt
     3721 ggtcaggaga tgtgctgccc catctgctgt agcgaatatg tgaaggggga ggtggcaact
     3781 gagctgccgt gccaccacta tttccacaag ccgtgtgtgt ccatctggct tcagaagtca
     3841 ggcacctgcc ccgtgtgccg ctgcatgttc cctcccccac tcctcgagga ctacaaagac
     3901 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga
     3961 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4021 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4081 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4141 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4201 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4261 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4741 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4801 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4861 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4921 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4981 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5041 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5281 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5881 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5941 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6001 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6061 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6181 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6241 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6301 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     7201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
