LOCUS       sequence-410377-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AraC-I113-LOV from a Trc promoter. For use in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4961)
  AUTHORS   Mathony J, Aschenbrenner S, Becker P, Niopek D
  TITLE     Dissecting the Determinants of Domain Insertion Tolerance and 
            Allostery in Proteins.
  JOURNAL   Adv Sci (Weinh). 2023 Oct;10(28):e2303496. doi: 
            10.1002/advs.202303496. Epub 2023 Aug 10.
  PUBMED    37562980
REFERENCE   2  (bases 1 to 4961)
  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: "Adv Sci 
            (Weinh)."; date: "2023-10"; volume: "10(28):e2303496. doi"; pages: "
            10.1002/advs.202303496. Epub 2023 Aug 10"
COMMENT     Sequence Label: pTrc_AraC-I113-LOV
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     167..184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(392..1474)
                     /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     1436..1455
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1475..1552)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(1597..1602,1603..1619,1620..1626)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1634..1650
                     /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)."
     CDS             2989..3006
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3024..3041)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3194..3280
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3372..3399
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3419..3513)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3534..4193)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LLSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4108..4127
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4194..4296)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      join(4386..4961,1..13)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4875..4894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
       61 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      181 cgaggaagcg gaagagcgcc acggtgattc attctgctaa ccagtaaggc aaccccgcca
      241 gcctagccgg gtcctcaacg acaggagcac gatcatgcta gtcatgcccc gcgcccaccg
      301 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
      361 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      481 gcgccagggt ggtttttctt ttcaccagtg acacgggcaa cagctgattg cccttcaccg
      541 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
      601 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
      661 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
      721 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgataccc tcattcagca
      781 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
      841 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
      901 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
      961 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     1021 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     1081 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     1141 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     1201 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     1261 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     1321 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     1381 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     1441 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     1501 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     1561 cgacgctctc ccttatgcga ctccgatccc gcaaatttga caattaatca tccggctcgt
     1621 ataatgtgtg gaattgtgag cggataacaa tttcacacaa ggagatatac catgtccgcg
     1681 gaagcgcaga acgatccgct gctgccgggc tatagcttta acgcgcatct ggtggcgggc
     1741 ctgaccccga ttgaagcgaa cggctatctg gattttttta ttgatcgccc gctgggcatg
     1801 aaaggctata ttctgaacct gaccattcgc ggccagggcg tggtgaaaaa ccagggccgc
     1861 gaatttgtgt gccgcccggg cgatattctg ctgtttccgc cgggcgaaat tcatcattat
     1921 ggccgccatc cggaagcgcg cgaatggtat catcagtggg tgtattttcg cccgcgcgcg
     1981 tattggcatg aatggctgaa ctggccgagc attagcggtt tagccacaac gctggaacgc
     2041 attgaaaaga atttcgtaat cacagacccg cgccttcccg acaatccaat tatttttgcg
     2101 tccgatagct tcctgcaatt aaccgaatac agccgcgaag aaattctggg tcgtaattgt
     2161 cgcttccttc aggggccaga gactgaccgt gctacggtac gcaaaatccg cgacgcaatc
     2221 gacaatcaaa cggaagtcac ggttcagttg attaactata cgaagagcgg aaaaaaattc
     2281 tggaatttat ttcacttgca gcctatgcgt gaccagaagg gcgatgtcca gtatttcatt
     2341 ggcgttcagc ttgatggtac cgagcatgtt cgcgatgctg cggagcgtga aggtgtaatg
     2401 ttaattaaaa agactgctga aaacattgat gaggcggcca aagggagctt tgcgaacacc
     2461 ggcttttttc gcccggatga agcgcatcag ccgcatttta gcgatctgtt tggccagatt
     2521 attaacgcgg gccagggcga aggccgctat agcgaactgc tggcgattaa cctgctggaa
     2581 cagctgctgc tgcgccgcat ggaagcgatt aacgaaagcc tgcatccgcc gatggataac
     2641 cgcgtgcgcg aagcgtgcca gtatattagc gatcatctgg cggatagcaa ctttgatatt
     2701 gcgagcgtgg cgcagcatgt gtgcctgagc ccgagccgcc tgagccatct gtttcgccag
     2761 cagctgggca ttagcgtgct gagctggcgc gaagatcagc gcattagcca ggcgaaactg
     2821 ctgctgagca ccacccgcat gccgattgcg accgtgggcc gcaacgtggg ctttgatgat
     2881 cagctgtatt ttagccgcgt gtttaaaaaa tgcaccggcg cgagcccgag cgaatttcgc
     2941 gcgggctgcg aagaaaaagt gaacgatgtg gcggtgaaac tgagcggtca ccatcaccat
     3001 caccattaag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     3061 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     3121 tcagaagtga aacgccgtag cgccgatggt agtgtggggt atccccatgc gagagtaggg
     3181 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3241 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3301 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3361 tcaaattaag cagaaggcca tcctgacgga tggccttttc tgaaaggagg aactatatat
     3421 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     3481 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
     3541 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     3601 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     3661 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     3721 aaatcaaaac tggtgaaact cacccaggga ttggctgaaa cgaaaaacat attctcaata
     3781 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     3841 tgtagaaact gccggaaatc gtcgtggtat tcactcaaga gcgatgaaaa cgtttcagtt
     3901 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     3961 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     4021 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     4081 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     4141 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     4201 tccttagctc ctgaaaatct cgataactaa aaaaatacgc ccggtagtga tcttatttca
     4261 ttatggtgaa agttggaacc tcttacgtgc cgatcaacag ttttattgtt catgaccaaa
     4321 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4381 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4441 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4501 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4561 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4621 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4681 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4741 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4801 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4861 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4921 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc g
//
