LOCUS       62056_6875        7167 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 7167)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             695..1411
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1460..2416
                     /codon_start=1
                     /label=TurboID
                     /note="mutant of the E. coli biotin ligase BioID engineered
                     for improved catalytic efficiency (Branon et al., 2018)"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2423..2449
                     /codon_start=1
                     /label=NES
                     /note="nuclear export signal from the HIV Rev protein
                     (Fischer et al., 1995)"
                     /translation="LPPLERLTL"
     promoter        complement(2486..2504)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2530..2754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2800..3228
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3242..3571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3638..4429
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4606..4739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4776..4792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4800..4816)
                     /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(4824..4854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4869..4890)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5178..5766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5940..6797)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6798..6902)
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcctg gaattctcgc caccatggtg agcaagggcg aggagctgtt
      721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      781 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      901 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1021 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1141 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1201 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1261 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1321 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1381 gatcactctc ggcatggacg agctgtacaa gggcaagccc atccccaacc ccctgctggg
     1441 cctggacagc accgctagca aagacaatac tgtgcctctg aagctgatcg ctctcctggc
     1501 taatggcgag ttccatagtg gcgaacagct gggagaaacc ctgggcatgt ccagggccgc
     1561 tatcaacaag cacattcaga ctctgcgcga ctggggcgtg gacgtgttca ccgtgcccgg
     1621 aaagggctac tctctgcccg agcctatccc gctgctgaac gctaaacaga ttctgggaca
     1681 gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc agtacctgct
     1741 ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc agcaggcagg
     1801 gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt acctgagtat
     1861 gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca tcggaattgt
     1921 catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat ggcccaatga
     1981 cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg gaataacagg
     2041 cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc gcgtggagga
     2101 aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc tggacaggaa
     2161 tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt tcgagcagga
     2221 aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca atagacccgt
     2281 gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg ataaacaggg
     2341 agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag aaatcagtct
     2401 gcggtctgcc gaaaagctgc agctgcctcc cctggagcgc ctgaccctgg actaatagct
     2461 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2521 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2581 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2641 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2701 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2761 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2821 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2881 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2941 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3001 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3061 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3121 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3181 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3601 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3661 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3721 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3781 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3841 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3901 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3961 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4021 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4081 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4141 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4201 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4261 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4321 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4381 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4441 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4501 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4561 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4741 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4801 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4861 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4921 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4981 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5041 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5101 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5161 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacaaac cccccgttca gcccgaccgc
     5461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5641 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5761 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5821 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5881 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5941 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6001 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6061 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6121 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6181 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6241 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6301 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6361 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6421 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6481 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6541 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6601 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6661 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6721 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6781 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6841 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6901 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6961 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7021 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7081 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     7141 gcgatgtacg ggccagatat acgcgtt
//
