LOCUS       62056_1590        6113 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             951..1646
                     /codon_start=1
                     /label=BioID2
                     /note="promiscuous R40G mutant of a biotin protein ligase
                     from Aquifex aeolicus (Kim et al., 2016)"
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             1647..1673
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    1709..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1979..2407
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2421..2751
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2818..3609
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3786..3919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3956..3972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3980..3996)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4004..4034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4049..4070)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4358..4946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5120..5977)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5978..6082)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttaaggcc tgttaaccgg tcgtacgtct ccggattcga attcggatcc ttcaagaacc
      961 tgatctggct gaaggaggtg gacagcaccc aggagagact gaaggagtgg aacgtgagct
     1021 acggcaccgc cctggtggcc gacagacaga ccaagggcag aggcggcctg ggcagaaagt
     1081 ggctgagcca ggagggcggc ctgtacttca gcttcctgct gaaccccaag gagttcgaga
     1141 acctgctgca gctgcccctg gtgctgggcc tgagcgtgag cgaggccctg gaggagatca
     1201 ccgagatccc cttcagcctg aagtggccca acgacgtgta cttccaggag aagaaggtga
     1261 gcggcgtgct gtgcgagctg agcaaggaca agctgatcgt gggcatcggc atcaacgtga
     1321 accagagaga gatccccgag gagatcaagg acagagccac caccctgtac gagatcaccg
     1381 gcaaggactg ggacagaaag gaggtgctgc tgaaggtgct gaagagaatc agcgagaacc
     1441 tgaagaagtt caaggagaag agcttcaagg agttcaaggg caagatcgag agcaagatgc
     1501 tgtacctggg cgaggaggtg aagctgctgg gcgagggcaa gatcaccggc aagctggtgg
     1561 gcctgagcga gaagggcggc gccctgatcc tgaccgagga gggcatcaag gagatcctga
     1621 gcggcgagtt cagcctgaga agaagctacc cttacgatgt accggattac gcatagaagc
     1681 ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1741 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1801 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1861 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1921 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2101 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2341 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2401 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2461 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2521 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2581 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2641 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2701 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2761 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2821 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2881 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2941 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3001 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3061 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3121 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3181 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3241 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3301 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3361 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3421 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3481 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3541 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3601 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3661 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3721 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3781 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3841 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3901 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3961 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4021 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4081 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4141 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4201 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4261 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4321 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4381 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4441 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4501 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     4561 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4621 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4681 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4741 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4801 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4861 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4921 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4981 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5041 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5101 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5161 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5221 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5281 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5341 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5401 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5461 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5521 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5581 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5641 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5701 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5761 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5821 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5881 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5941 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6001 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6061 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
