LOCUS       V036766                 7649 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036766
VERSION     V036766
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             941..2167
                     /label="CARNMT1(NM_152420)"
                     /note="CARNMT1(NM_152420)"
                     /gene="CARNMT1"
     CDS             2180..3139
                     /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             3146..3211
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3249..3473
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3519..3947
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3961..4290
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4357..5151
                     /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    5325..5446
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5495..5511)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5519..5535
                     /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(5543..5573)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5588..5609
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5897..6482)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6653..7513)
                     /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(7514..7618)
                     /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 atccgccacc atgcagcgac ggcgtcgccc
      961 tccgccgccc acctcccggc tgcccgaggg ctgcggggga ggaggcggtg gcagcgagga
     1021 ggtggaagtg cagttttccg ccgggcgttg gggctcggcc gcggcggttt cggcggcagc
     1081 ggcggcggcc acgcgcagca ccgaggagga ggaggagagg cttgagcgtg agcacttctg
     1141 gaagatcatt aatgccttcc gctactacgg caccagtatg catgagcggg tgaaccgaac
     1201 agaaagacag tttcgatcac ttccagctaa ccaacagaaa ctacttcctc agtttcttct
     1261 tcacttggac aagatccgga aatgcattga tcataatcaa gaaatactac tgaccattgt
     1321 gaatgattgc atacatatgt ttgaaaataa agaatatgga gaagatggga atggaaagat
     1381 tatgccagca tctacatttg acatggataa gttaaaatcc acgctgaaac agtttgtgag
     1441 agactggagt gaaactggga aagcagaaag ggatgcctgt taccagccaa tcattaaaga
     1501 aattttaaaa aattttccaa aagagagatg ggatccttct aaagtaaata ttctggtacc
     1561 tggtgctgga ctaggaagac tggcctggga aatagctatg ctaggttatg cttgtcaagg
     1621 aaatgaatgg agttttttta tgctcttttc ttccaacttt gtactcaaca gatgttctga
     1681 aattaataaa tataaacttt atccttggat ccatcagttt agcaataacc ggagatcagc
     1741 tgatcagatt cgacccatct ttttccctga tgttgacccc cacagtcttc ctcctggttc
     1801 taacttttct atgacagcag gagattttca agagatttat tcagaatgca atacctggga
     1861 ctgtattgct acctgtttct tcatagacac agctcacaat gtaattgatt atattgatac
     1921 aatatggaaa atactcaagc caggtggaat ttggataaat ctaggtcctc tgctgtacca
     1981 ctttgaaaat ctggcaaatg aactttccat agaattgagc tatgaggata taaaaaacgt
     2041 tgttctgcag tatggattca aggtagaggt ggaaaaagaa tctgtattgt caacatatac
     2101 tgtgaatgat ctctctatga tgaaatacta ctatgaatgt gtcttgtttg tggtccgtaa
     2161 gccacaagaa ttcgccacca tgaaagacaa tactgtgcct ctgaagctga tcgctctcct
     2221 ggctaatggc gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc
     2281 cgctatcaac aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc
     2341 cggaaagggc tactctctgc ccgagcctat cccgctgctg aacgctaaac agattctggg
     2401 acagctggac ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct
     2461 gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc
     2521 agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag
     2581 tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat
     2641 tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa
     2701 tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac
     2761 aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga
     2821 ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag
     2881 gaatactctg gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca
     2941 ggaaggcctg gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc
     3001 cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca
     3061 gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag
     3121 tctgcggtct gccgaaaagc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3181 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     3241 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3301 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3361 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3421 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3481 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3541 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3601 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3661 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3721 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3781 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3841 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3901 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3961 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4021 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4081 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4141 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4201 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4261 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4321 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4381 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4441 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4501 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4561 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4621 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4681 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4741 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4801 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4861 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4921 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4981 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5041 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5101 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5161 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5221 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5281 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5341 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5401 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5461 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5641 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5701 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6301 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6421 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6601 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6661 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6721 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6781 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6841 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6901 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6961 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7021 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7081 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7141 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7201 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7261 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7321 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7381 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7441 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7501 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7561 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7621 catttccccg aaaagtgcca cctgacgtc
//
