LOCUS       V033554                 8429 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033554
VERSION     V033554
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             665..1624
                     /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             1631..3085
                     /label="RCOR1(NM_015156)"
                     /note="RCOR1(NM_015156)"
                     /gene="RCOR1"
     CDS             3092..3181
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3205..3793
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3859..3907
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4109..4537
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4551..4880
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4947..5741
                     /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    5917..6038
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6087..6103)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6111..6127
                     /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(6135..6165)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6180..6201
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6489..7077)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7248..8108)
                     /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(8109..8213)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgc
      661 caccatgaaa gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt
      721 ccatagtggc gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca
      781 cattcagact ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc
      841 tctgcccgag cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg
      901 gagcgtggca gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg
      961 cgagctgaag agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag
     1021 agggaggaaa tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct
     1081 gaagcgggga ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc
     1141 gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca
     1201 ggatagaaag ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca
     1261 gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa
     1321 tcagggctgg atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc
     1381 tacgctgatc cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc
     1441 atatctgcca cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat
     1501 tggggacaaa gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct
     1561 ggaacaggac ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga
     1621 aaagggtacc atgccggcca tggtggagaa gggccccgag gtctcaggga agcggagagg
     1681 gaggaacaac gcggccgcct ccgcctccgc cgccgccgcc tccgccgccg cctcggccgc
     1741 ctgcgcctcg ccagccgcca ctgccgcctc gggcgccgcc gcctcctcag cctcggccgc
     1801 cgccgcctca gccgccgccg cccccaataa tggccagaat aaaagtttgg cggcggcggc
     1861 gcccaatggc aacagcagca gcaactcctg ggaggaaggc agctcgggct cgtccagcga
     1921 cgaggagcac ggtggcggtg gcatgagggt cggaccccag taccaggcgg tggtgcccga
     1981 cttcgacccc gccaaactgg caagacgcag tcaagaacgg gacaatcttg gcatgttggt
     2041 ctggtcaccc aatcaaaatc tgtcagaagc aaagttggat gaatacattg ccattgccaa
     2101 agaaaagcat gggtacaaca tggaacaggc tcttgggatg ctcttctggc ataaacataa
     2161 tatcgaaaag tcattggctg atttgcccaa ctttacccct ttcccagatg agtggactgt
     2221 ggaagataaa gtcttatttg agcaagcctt tagttttcat gggaaaactt ttcatagaat
     2281 ccaacaaatg cttccagata aatctatagc aagtctggtg aaattttact attcttggaa
     2341 gaagacgagg actaaaacta gtgtgatgga tcgccatgcc cggaaacaaa aacgggagcg
     2401 ggaggagagc gaggatgaac tggaagaggc aaatggaaac aatcccattg acattgaggt
     2461 tgatcaaaac aaggaaagca aaaaggaggt tccccctact gagacagttc ctcaggtcaa
     2521 aaaagaaaaa catagcacac aagctaaaaa tagagcaaaa aggaaacctc caaaaggaat
     2581 gtttctttct caagaagatg tggaggctgt ttctgccaat gccactgctg ctaccacggt
     2641 gctgagacaa ctagacatgg aattggtttc agtcaaacga cagatccaga atattaaaca
     2701 gacaaacagt gctctcaaag aaaaacttga tggtggaata gaaccatatc gacttccaga
     2761 ggtcattcag aaatgtaatg cacgttggac tacagaagag cagcttctcg ccgtacaagc
     2821 catcaggaaa tatggccgag attttcaggc aatctcagac gtgattggga acaaatcagt
     2881 ggtacaagtg aaaaactttt ttgtaaatta tcgacgccgc ttcaacatag atgaagtttt
     2941 acaagaatgg gaggcagaac atggtaaaga agagaccaat gggcccagta accagaagcc
     3001 tgtgaagtcc ccagataatt ccattaagat gcccgaagag gaagacgagg ctcctgttct
     3061 ggatgtcaga tatgcatctg cctccctcga ggaacaaaaa ctaatatcag aggaagattt
     3121 ggaacagaaa ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt
     3181 atagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     3241 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3301 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3361 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3421 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3481 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3541 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     3601 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3661 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3721 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3781 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     3841 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3901 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3961 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4021 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     4081 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4141 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4201 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4261 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4321 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4381 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4441 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     4501 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4561 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4621 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4681 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4741 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4801 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4861 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4921 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4981 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5041 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5101 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5161 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5221 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5281 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5341 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5401 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5461 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5521 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5581 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5641 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5701 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     5761 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5821 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5881 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5941 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6001 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6061 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6121 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6181 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6241 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6301 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6661 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6901 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8221 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     8281 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     8341 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     8401 ccgacaattg catgaagaat ctgcttagg
//
