LOCUS       V044267                 7265 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044267
VERSION     V044267
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..1753
                     /label="TARM1(NM_001135686)"
                     /note="TARM1(NM_001135686)"
                     /gene="TARM1"
     CDS             1796..2755
                     /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             2762..2827
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2865..3089
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3135..3563
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3577..3906
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3973..4767
                     /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    4941..5062
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5111..5127)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5135..5151
                     /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(5159..5189)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5204..5225
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5513..6098)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6269..7129)
                     /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(7130..7234)
                     /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 atgatcccta agctgctttc
      961 cctcctctgt ttcagactgt gcgtgggcca aggagacaca aggggagatg ggtcactgcc
     1021 caagccgtcc ctcagtgcct ggcccagctc ggtggtccct gccaacagca atgtgacgct
     1081 gcgatgttgg actcctgcca gaggtgtgag ctttgttctc aggaagggag gaattattct
     1141 ggagtccccg aagccccttg attctacaga gggcgcggcc gaatttcacc tcaataatct
     1201 aaaagtcaga aatgctggag agtacacctg tgaatactac agaaaagcat ccccccacat
     1261 cctttcacag cacagtgacg tccttctact gttggtgaca ggacatttat ctaaaccttt
     1321 cctccgaacc taccaaaggg gtacagtgac cgcaggtgga agggtgactc tgcagtgcca
     1381 gaagcgagac caattgtttg tgcctatcat gttcgctcta ctgaaggcag ggacgccatc
     1441 acccatccag ctgcagagtc cagcggggaa ggagatagac ttctctctgg tggacgtgac
     1501 agccggcgat gctgggaact acagctgcat gtactaccag acaaagtctc ccttctgggc
     1561 ctcagaaccc agtgatcagc ttgagatatt ggtgacagtt cccccaggta ccacatcgag
     1621 caactactcc ctgggtaact tcgtacgact gggtctggct gccgtaattg tggttatcat
     1681 gggagctttc ctggtggagg cctggtacag ccggaatgtg tctccaggtg aatcagaggc
     1741 cttcaaacca gagggtggag gcggatcagg aggcggtgga tctgaattcg ccaccatgaa
     1801 agacaatact gtgcctctga agctgatcgc tctcctggct aatggcgagt tccatagtgg
     1861 cgaacagctg ggagaaaccc tgggcatgtc cagggccgct atcaacaagc acattcagac
     1921 tctgcgcgac tggggcgtgg acgtgttcac cgtgcccgga aagggctact ctctgcccga
     1981 gcctatcccg ctgctgaacg ctaaacagat tctgggacag ctggacggcg ggagcgtggc
     2041 agtcctgcct gtggtcgact ccaccaatca gtacctgctg gatcgaatcg gcgagctgaa
     2101 gagtggggat gcttgcattg cagaatatca gcaggcaggg agaggaagca gagggaggaa
     2161 atggttctct ccttttggag ctaacctgta cctgagtatg ttttggcgcc tgaagcgggg
     2221 accagcagca atcggcctgg gcccggtcat cggaattgtc atggcagaag cgctgcgaaa
     2281 gctgggagca gacaaggtgc gagtcaaatg gcccaatgac ctgtatctgc aggatagaaa
     2341 gctggcaggc atcctggtgg agctggccgg aataacaggc gatgctgcac agatcgtcat
     2401 tggcgccggg attaacgtgg ctatgaggcg cgtggaggaa agcgtggtca atcagggctg
     2461 gatcacactg caggaagcag ggattaacct ggacaggaat actctggccg ctacgctgat
     2521 ccgagagctg cgggcagccc tggaactgtt cgagcaggaa ggcctggctc catatctgcc
     2581 acggtgggag aagctggata acttcatcaa tagacccgtg aagctgatca ttggggacaa
     2641 agagattttc gggattagcc gggggattga taaacaggga gccctgctgc tggaacagga
     2701 cggagttatc aaaccctgga tgggcggaga aatcagtctg cggtctgccg aaaagctcga
     2761 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2821 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2881 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2941 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3001 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3061 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3121 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3181 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3241 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3301 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3361 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3421 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3481 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3541 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3781 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3841 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3901 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3961 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4021 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4081 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4141 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4201 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4261 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4321 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4381 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4441 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4501 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4561 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4621 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4681 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4741 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4801 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4861 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4921 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4981 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5041 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5101 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5161 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5221 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5281 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5341 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5401 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5941 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgtc
//
