LOCUS       sequence-410371-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AraC-S170-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-S170-LOV
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(388..1470)
                     /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     1432..1451
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1471..1548)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(1593..1598,1599..1615,1616..1622)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1630..1646
                     /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             2985..3002
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3020..3037)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3190..3276
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3368..3395
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3415..3509)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3530..4189)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LLSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4104..4123
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4190..4292)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      join(4382..4961,1..9)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4871..4890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
       61 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      181 gaagcggaag agcgccacgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
      241 agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag
      301 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
      361 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
      421 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      481 cagggtggtt tttcttttca ccagtgacac gggcaacagc tgattgccct tcaccgcctg
      541 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
      601 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
      661 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
      721 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg ataccctcat tcagcatttg
      781 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
      841 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
      901 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
      961 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     1021 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     1081 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     1141 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     1201 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     1261 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     1321 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     1381 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     1441 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     1501 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     1561 gctctccctt atgcgactcc gatcccgcaa atttgacaat taatcatccg gctcgtataa
     1621 tgtgtggaat tgtgagcgga taacaatttc acacaaggag atataccatg tccgcggaag
     1681 cgcagaacga tccgctgctg ccgggctata gctttaacgc gcatctggtg gcgggcctga
     1741 ccccgattga agcgaacggc tatctggatt tttttattga tcgcccgctg ggcatgaaag
     1801 gctatattct gaacctgacc attcgcggcc agggcgtggt gaaaaaccag ggccgcgaat
     1861 ttgtgtgccg cccgggcgat attctgctgt ttccgccggg cgaaattcat cattatggcc
     1921 gccatccgga agcgcgcgaa tggtatcatc agtgggtgta ttttcgcccg cgcgcgtatt
     1981 ggcatgaatg gctgaactgg ccgagcattt ttgcgaacac cggctttttt cgcccggatg
     2041 aagcgcatca gccgcatttt agcgatctgt ttggccagat tattaacgcg ggccagggcg
     2101 aaggccgcta tagcgaactg ctggcgatta acctgctgga acagctgctg ctgcgccgca
     2161 tggaagcgat taacgaaagc agcggtttag ccacaacgct ggaacgcatt gaaaagaatt
     2221 tcgtaatcac agacccgcgc cttcccgaca atccaattat ttttgcgtcc gatagcttcc
     2281 tgcaattaac cgaatacagc cgcgaagaaa ttctgggtcg taattgtcgc ttccttcagg
     2341 ggccagagac tgaccgtgct acggtacgca aaatccgcga cgcaatcgac aatcaaacgg
     2401 aagtcacggt tcagttgatt aactatacga agagcggaaa aaaattctgg aatttatttc
     2461 acttgcagcc tatgcgtgac cagaagggcg atgtccagta tttcattggc gttcagcttg
     2521 atggtaccga gcatgttcgc gatgctgcgg agcgtgaagg tgtaatgtta attaaaaaga
     2581 ctgctgaaaa cattgatgag gcggccaaag ggagcctgca tccgccgatg gataaccgcg
     2641 tgcgcgaagc gtgccagtat attagcgatc atctggcgga tagcaacttt gatattgcga
     2701 gcgtggcgca gcatgtgtgc ctgagcccga gccgcctgag ccatctgttt cgccagcagc
     2761 tgggcattag cgtgctgagc tggcgcgaag atcagcgcat tagccaggcg aaactgctgc
     2821 tgagcaccac ccgcatgccg attgcgaccg tgggccgcaa cgtgggcttt gatgatcagc
     2881 tgtattttag ccgcgtgttt aaaaaatgca ccggcgcgag cccgagcgaa tttcgcgcgg
     2941 gctgcgaaga aaaagtgaac gatgtggcgg tgaaactgag cggtcaccat caccatcacc
     3001 attaagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     3061 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     3121 aagtgaaacg ccgtagcgcc gatggtagtg tggggtatcc ccatgcgaga gtagggaact
     3181 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3241 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     3301 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     3361 attaagcaga aggccatcct gacggatggc cttttctgaa aggaggaact atatattctc
     3421 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     3481 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     3541 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     3601 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     3661 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     3721 caaaactggt gaaactcacc cagggattgg ctgaaacgaa aaacatattc tcaataaacc
     3781 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     3841 gaaactgccg gaaatcgtcg tggtattcac tcaagagcga tgaaaacgtt tcagtttgct
     3901 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     3961 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     4021 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     4081 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     4141 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     4201 tagctcctga aaatctcgat aactaaaaaa atacgcccgg tagtgatctt atttcattat
     4261 ggtgaaagtt ggaacctctt acgtgccgat caacagtttt attgttcatg accaaaatcc
     4321 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4381 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4441 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4501 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4561 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4621 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4681 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4741 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4801 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4861 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4921 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag c
//
