LOCUS       V046419                 7403 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046419
VERSION     V046419
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..1876
                     /label="AIMP1(NM_004757)"
                     /note="AIMP1(NM_004757)"
                     /gene="AIMP1"
     CDS             1934..2893
                     /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             2900..2965
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3003..3227
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3273..3701
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3715..4044
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4111..4905
                     /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    5079..5200
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5249..5265)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5273..5289
                     /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(5297..5327)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5342..5363
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5651..6236)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6407..7267)
                     /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(7268..7372)
                     /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 atggcaaata atgatgctgt
      961 tctgaagaga ctggagcaga agggtgcaga ggcagatcaa atcattgaat atcttaagca
     1021 gcaagtttct ctacttaagg agaaagcaat tttgcaggca actttgaggg aagagaagaa
     1081 acttcgagtt gaaaatgcta aactgaagaa agaaattgaa gaactgaaac aagagctaat
     1141 tcaggcagaa attcaaaatg gagtgaagca aataccattt ccatctggta ctccactgca
     1201 cgctaattct atggtttctg aaaatgtgat acagtctaca gcagtaacaa ccgtatcttc
     1261 tggtaccaaa gaacagataa aaggaggaac aggagacgaa aagaaagcga aagagaaaat
     1321 tgaaaagaaa ggagagaaga aggagaaaaa acagcaatca atagctggaa gtgccgactc
     1381 taagccaata gatgtttccc gtctggatct tcgaattggt tgcatcataa ctgctagaaa
     1441 acaccctgat gcagattctt tgtatgtgga agaagtagat gtcggagaaa tagccccaag
     1501 gacagttgtc agtggcctgg tgaatcatgt tcctcttgaa cagatgcaaa atcggatggt
     1561 gattttactt tgtaacctga aacctgcaaa gatgagggga gtattatctc aagcaatggt
     1621 catgtgtgct agttcaccag agaaaattga aatcttggct cctccaaatg ggtctgttcc
     1681 tggagacaga attacttttg atgctttccc aggagagcct gacaaggagc tgaatcctaa
     1741 gaagaagatt tgggagcaga tccagcctga tcttcacact aatgatgagt gtgtggctac
     1801 atacaaagga gttccctttg aggtgaaagg gaagggagta tgtagggctc aaaccatgag
     1861 caacagtgga atcaaaggag gcggaggtag tggaggaggg ggaagcggag gcggagggag
     1921 tgaattcgcc accatgaaag acaatactgt gcctctgaag ctgatcgctc tcctggctaa
     1981 tggcgagttc catagtggcg aacagctggg agaaaccctg ggcatgtcca gggccgctat
     2041 caacaagcac attcagactc tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa
     2101 gggctactct ctgcccgagc ctatcccgct gctgaacgct aaacagattc tgggacagct
     2161 ggacggcggg agcgtggcag tcctgcctgt ggtcgactcc accaatcagt acctgctgga
     2221 tcgaatcggc gagctgaaga gtggggatgc ttgcattgca gaatatcagc aggcagggag
     2281 aggaagcaga gggaggaaat ggttctctcc ttttggagct aacctgtacc tgagtatgtt
     2341 ttggcgcctg aagcggggac cagcagcaat cggcctgggc ccggtcatcg gaattgtcat
     2401 ggcagaagcg ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc ccaatgacct
     2461 gtatctgcag gatagaaagc tggcaggcat cctggtggag ctggccggaa taacaggcga
     2521 tgctgcacag atcgtcattg gcgccgggat taacgtggct atgaggcgcg tggaggaaag
     2581 cgtggtcaat cagggctgga tcacactgca ggaagcaggg attaacctgg acaggaatac
     2641 tctggccgct acgctgatcc gagagctgcg ggcagccctg gaactgttcg agcaggaagg
     2701 cctggctcca tatctgccac ggtgggagaa gctggataac ttcatcaata gacccgtgaa
     2761 gctgatcatt ggggacaaag agattttcgg gattagccgg gggattgata aacagggagc
     2821 cctgctgctg gaacaggacg gagttatcaa accctggatg ggcggagaaa tcagtctgcg
     2881 gtctgccgaa aagctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     2941 cgactacaag gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc
     3001 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3061 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3121 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3181 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3241 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3481 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3601 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3661 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4081 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4141 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4201 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4261 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4321 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4381 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4441 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4501 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4561 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4621 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4681 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4741 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4801 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4861 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4921 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5041 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5221 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5281 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5341 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5401 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5461 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5821 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6061 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6181 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6241 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6421 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6481 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6541 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6601 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7321 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7381 cccgaaaagt gccacctgac gtc
//
