LOCUS       sequence-418280-        8441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible promoter (lacUV5) expressing AcrA and 
            AcrB-AsLOV2*(543). AsLOV2*(543) is also described in the literature 
            as LOVdeg..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8441)
  AUTHORS   Tague N, Coriano-Ortiz C, Sheets MB, Dunlop MJ
  TITLE     Light-inducible protein degradation in E. coli with the LOVdeg tag.
  JOURNAL   Elife. 2024 Jan 25;12:RP87303. doi: 10.7554/eLife.87303.
  PUBMED    38270583
REFERENCE   2  (bases 1 to 8441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2024-01-25"; volume: "12:RP87303. doi"; pages: " 
            10.7554/eLife.87303"
COMMENT     Sequence Label: pBbA5k-AcrAB-AsLOV2*(543)
FEATURES             Location/Qualifiers
     source          1..8441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(424..518)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(549..1343)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(660..679)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1270..1289
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1528..2610)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2572..2591
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2611..2688)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2894..2913
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(2999..3004,3005..3022,3023..3029)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3037..3053
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     terminator      7930..8001
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8017..8044
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(8072..8089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8206..8441,1..310))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(8286..8305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
       61 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
      121 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
      181 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
      241 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
      301 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
      361 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct
      421 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
      481 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag
      541 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
      601 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
      661 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
      721 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
      781 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
      841 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
      901 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
      961 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     1021 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     1081 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     1141 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     1201 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     1261 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     1321 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat
     1381 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     1441 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcggtgc ctaatgagtg
     1501 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1561 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1621 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     1681 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     1741 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     1801 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     1861 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     1921 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     1981 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     2041 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     2101 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     2161 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     2221 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     2281 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     2341 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     2401 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     2461 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     2521 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     2581 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     2641 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     2701 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2761 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2821 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2881 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2941 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgtttaggca ccccaggctt
     3001 tacactttat gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaga
     3061 attcaaaaga tcttttaaga aggagatata catatgaaca aaaacagagg gtttacgcct
     3121 ctggcggtcg ttctgatgct ctcaggcagc ttagccctaa caggatgtga cgacaaacag
     3181 gcccaacaag gtggccagca gatgcccgcc gttggcgtag taacagtcaa aactgaacct
     3241 ctgcagatca caaccgagct tccgggtcgc accagtgcct accggatcgc agaagttcgt
     3301 cctcaagtta gcgggattat cctgaagcgt aatttcaaag aaggtagcga catcgaagca
     3361 ggtgtctctc tctatcagat tgatcctgcg acctatcagg cgacatacga cagtgcgaaa
     3421 ggtgatctgg cgaaagccca ggctgcagcc aatatcgcgc aattgacggt gaatcgttat
     3481 cagaaactgc tcggtactca gtacatcagt aagcaagagt acgatcaggc tctggctgat
     3541 gcgcaacagg cgaatgctgc ggtaactgcg gcgaaagctg ccgttgaaac tgcgcggatc
     3601 aatctggctt acaccaaagt cacctctccg attagcggtc gcattggtaa gtcgaacgtg
     3661 acggaaggcg cattggtaca gaacggtcag gcgactgcgc tggcaaccgt gcagcaactt
     3721 gatccgatct acgttgatgt gacccagtcc agcaacgact tcctgcgcct gaaacaggaa
     3781 ctggcgaatg gcacgctgaa acaagagaac ggcaaagcca aagtgtcact gatcaccagt
     3841 gacggcatta agttcccgca ggacggtacg ctggaattct ctgacgttac cgttgatcag
     3901 accactgggt ctatcaccct acgcgctatc ttcccgaacc cggatcacac tctgctgccg
     3961 ggtatgttcg tgcgcgcacg tctggaagaa gggcttaatc caaacgctat tttagtcccg
     4021 caacagggcg taacccgtac gccgcgtggc gatgccaccg tactggtagt tggcgcggat
     4081 gacaaagtgg aaacccgtcc gatcgttgca agccaggcta ttggcgataa gtggctggtg
     4141 acagaaggtc tgaaagcagg cgatcgcgta gtaataagtg ggctgcagaa agtgcgtcct
     4201 ggtgtccagg taaaagcaca agaagttacc gctgataata accagcaagc cgcaagcggt
     4261 gctcagcctg aacagtccaa gtcttaactt aaacaggagc cgttaagaca tgcctaattt
     4321 ctttatcgat cgcccgattt ttgcgtgggt aatcgccatt atcatcatgt tggcaggggg
     4381 gctggcgatc ctcaaactgc cggtggcgca atatcctacg attgcaccgc cggcagtaac
     4441 gatctccgcc tcctaccccg gcgctgatgc gaaaacagtg caggacacgg tgacacaggt
     4501 tatcgaacag aatatgaacg gtatcgataa cctgatgtac atgtcctcta acagtgactc
     4561 cacgggtacc gtgcagatca ccctgacctt tgagtctggt actgatgcgg atatcgcgca
     4621 ggttcaggtg cagaacaaac tgcagctggc gatgccgttg ctgccgcaag aagttcagca
     4681 gcaaggggtg agcgttgaga aatcatccag cagcttcctg atggttgtcg gcgttatcaa
     4741 caccgatggc accatgacgc aggaggatat ctccgactac gtggcggcga atatgaaaga
     4801 tgccatcagc cgtacgtcgg gcgtgggtga tgttcagttg ttcggttcac agtacgcgat
     4861 gcgtatctgg atgaacccga atgagctgaa caaattccag ctaacgccgg ttgatgtcat
     4921 taccgccatc aaagcgcaga acgcccaggt tgcggcgggt cagctcggtg gtacgccgcc
     4981 ggtgaaaggc caacagctta acgcctctat tattgctcag acgcgtctga cctctactga
     5041 agagttcggc aaaatcctgc tgaaagtgaa tcaggatggt tcccgcgtgc tgctgcgtga
     5101 cgtcgcgaag attgagctgg gtggtgagaa ctacgacatc atcgcagagt ttaacggcca
     5161 accggcttcc ggtctgggga tcaagctggc gaccggtgca aacgcgctgg ataccgctgc
     5221 ggcaatccgt gctgaactgg cgaagatgga accgttcttc ccgtcgggtc tgaaaattgt
     5281 ttacccatac gacaccacgc cgttcgtgaa aatctctatt cacgaagtgg ttaaaacgct
     5341 ggtcgaagcg atcatcctcg tgttcctggt tatgtatctg ttcctgcaga acttccgcgc
     5401 gacgttgatt ccgaccattg ccgtaccggt ggtattgctc gggacctttg ccgtccttgc
     5461 cgcctttggc ttctcgataa acacgctaac aatgttcggg atggtgctcg ccatcggcct
     5521 gttggtggat gacgccatcg ttgtggtaga aaacgttgag cgtgttatgg cggaagaagg
     5581 tttgccgcca aaagaagcta cccgtaagtc gatggggcag attcagggcg ctctggtcgg
     5641 tatcgcgatg gtactgtcgg cggtattcgt accgatggcc ttctttggcg gttctactgg
     5701 tgctatctat cgtcagttct ctattaccat tgtttcagca atggcgctgt cggtactggt
     5761 ggcgttgatc ctgactccag ctctttgtgc caccatgctg aaaccgattg ccaaaggcga
     5821 tcacggggaa ggtaaaaaag gcttcttcgg ctggtttaac cgcatgttcg agaagagcac
     5881 gcaccactac accgacagcg taggcggtat tctgcgcagt acggggcgtt acctggtgct
     5941 gtatctgatc atcgtggtcg gcatggccta tctgttcgtg cgtctgccaa gctccttctt
     6001 gccagatgag gaccagggcg tgtttatgac catggttcag ctgccagcag gtgcaacgca
     6061 ggaacgtaca cagaaagtgc tcaatgaggt aacgcattac tatctgacca aagaaaagaa
     6121 caacgttgag tcggtgttcg ccgttaacgg cttcggcttt gcgggacgtg gtcagaatac
     6181 cggtattgcg ttcgtttcct tgaaggactg ggccgatcgt ccgggcgaag aaaacaaagt
     6241 tgaagcgatt accatgcgtg caacacgcgc tttctcgcaa atcaaagatg cgatggtttt
     6301 cgcctttaac ctgcccgcaa tcgtggaact gggtactgca accggctttg actttgagct
     6361 gattgaccag gctggccttg gtcacgaaaa actgactcag gcgcgtaacc agttgcttgc
     6421 agaagcagcg aagcaccctg atatgttgac cagcgtacgt ccaaacggtc tggaagatac
     6481 cccgcagttt aagattgata tcgaccagga aaaagcgcag gcgctgggtg tttctatcaa
     6541 cgacattaac accactctgg gcgctgcatg gggcggcagc tatgtgaacg actttatcga
     6601 ccgcggtcgt gtgaagaaag tttatgtcat gtcagaagcg aaataccgta tgctgccgga
     6661 tgatatcggc gactggtatg ttcgtgctgc tgatggtcag atggtgccat tctcggcgtt
     6721 ctcctcttct cgttgggagt acggttcgcc gcgtctggaa cgttacaacg gcctgccatc
     6781 catggaaatc ttaggccagg cggcaccggg taaaagtacc ggtgaagcaa tggagctgat
     6841 ggaacaactg gcgagcaaac tgcctaccgg tgttggctat gactggacgg ggatgtccta
     6901 tcaggaacgt ctctccggca accaggcacc ttcactgtac gcgatttcgt tgattgtcgt
     6961 gttcctgtgt ctggcggcgc tgtacgagag ctggtcgatt ccgttctccg ttatgctggt
     7021 cgttccgctg ggggttatcg gtgcgttgct ggctgccacc ttccgtggcc tgaccaatga
     7081 cgtttacttc caggtaggcc tgctcacaac cattgggttg tcggcgaaga acgcgatcct
     7141 tatcgtcgaa ttcgccaaag acttgatgga taaagaaggt aaaggtctga ttgaagcgac
     7201 gcttgatgcg gtgcggatgc gtttacgtcc gatcctgatg acctcgctgg cgtttatcct
     7261 cggcgttatg ccgctggtta tcagtactgg tgctggttcc ggcgcgcaga acgcagtagg
     7321 taccggtgta atgggcggga tggtgaccgc aacggtactg gcaatcttct tcgttccggt
     7381 attctttgtg gtggttcgcc gccgctttag ccgcaagaat gaagatatcg agcacagcca
     7441 tactgtcgat catcattcga gcggatcagg tttggctact acacttgaac gtattgagaa
     7501 gaactttgtc attactgacc caagattgcc agataatccc attatattcg cgtccgatag
     7561 tttcttgcag ttgacagaat atagccgtga agaaattttg ggaagaaact gtcgcttctt
     7621 acaaggcccg gaaaccgatc gcgcaaccgt ccgtaagatt cgcgacgcca tcgataatca
     7681 aaccgaggtt acggtgcaat taattaacta cacgaaatcc ggtaagaagt tttggaatgt
     7741 atttcatttg caacccatgc gtgaccagaa aggagatgta caatatttca tcggagttca
     7801 actcgacggt acggagcgcc ttcgcggggc agcggaacgc gaagccgtta tgttgattaa
     7861 gaaaaccgcg ttccaaatcg cggaggcggc ataatgagga tccaaactcg agtaaggatc
     7921 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     7981 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     8041 tgcgtttata cctagggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     8101 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     8161 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     8221 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     8281 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     8341 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     8401 cactcagttc cgggtaggca gttcgctcca agctggactg t
//
