LOCUS       sequence-484660-        4969 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  Expression Plasmid containing a LacI repressed sd2 AraC R2 PANCE 
            cassette with a LOV domain insertion at S170.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 4969)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDest_PANCEd_AraC_S170_LOV_sd2_R2_C1
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(379..1461)
                     /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     1423..1442
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1462..1539)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(1584..1589,1590..1606,1607..1613)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1621..1637
                     /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)."
     primer_bind     complement(3019..3036)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3189..3275
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3367..3394
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3414..3508)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3529..4188)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LLSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4103..4122
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4189..4291)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4381..4969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4870..4889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgccacg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
      241 ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa ggagctgact
      301 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
      361 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      421 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
      481 ttttcttttc accagtgaca cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
      541 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
      601 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      661 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
      721 atcgttggca accagcatcg cagtgggaac gataccctca ttcagcattt gcatggtttg
      781 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
      841 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
      901 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
      961 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     1021 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     1081 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     1141 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     1201 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     1261 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     1321 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     1381 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     1441 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     1501 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     1561 tatgcgactc cgatcccgca aatttgacaa ttaatcatcc ggctcgtata atgtgtggaa
     1621 ttgtgagcgg ataacaattg gagtaaaaaa aaaggaaaat gtccgcgaaa gcgcagaacg
     1681 atccgctgct gccgggctat agctttaacg cacatctggt ggcgggcctg accccgattg
     1741 aagcgaacgg ctatctggat ttttttattg atcgcccgct gggcatgaaa ggctatattc
     1801 tgaacctgac cattcgcggc cagggcgtgg tgaaaaacca gggccgcgaa tttgtgtgcc
     1861 gcccgggcga tattctgctg tttccgccgg gcgaaattca tcattatggc cgccatccgg
     1921 aagcgcgaga atggtatcat cagtgggtgt attttcgccc gcgcgcgtat tggcatgaat
     1981 ggctgaactg gccgagcatt tttgcgaaca ccggcttttt tcgcccggat gaagcgcatc
     2041 agccgcattt tagcgatctg tttggccaga ttattaacgc gggccagggc gaaggcctct
     2101 atagcgaact gctgacaatt aacctgctgg aacagctgct gctgcgccgc atggaagcga
     2161 ttaacgaaag cagcggttta gccacaacgc tggaacgcat tgaaaagaat ttcgtaatca
     2221 cagacccgcg ccttcccgac aatccaatta tttttgcgtc cgatagcttc ctgcaattaa
     2281 ccgaatacag ccgcgaagaa attctgggtc gtaattgtcg cttccttcag gggccagaga
     2341 ctgaccgtgc tacggtacgc aaaatccgcg acgcaatcga caatcaaacg gaagtcacgg
     2401 ttcagttgat taactatacg aagagcggaa aaaaattctg gaatttattt cacttgcagc
     2461 ctatgcgtga ccagaagggc gatgtccagt atttcattgg cgttcagctt gatggtaccg
     2521 agcatgttcg cgatgctgcg gagcgtgaag gtgtaatgtt aattaaaaag actgctgaaa
     2581 acattgatga ggcggccaaa gggagcctgc atccgccgat ggataaccgc gtgcgcgaag
     2641 cgtgccagta tattagcgat catctggcgg atagcaactt tgatattgcg agcgtggcgc
     2701 agcatgtgtg cctgagccct agccgcctga gccatctgtt tcgccagcag ctgggcatta
     2761 gcgtgctgag ctggcgcgaa gatcagcgca ttagccaggc gaaactgctg ctgagcacca
     2821 cccgcatgcc gattgcgacc gtgggcctca acgtgggctt tgatgatcag ctgtatttta
     2881 gccgcgtgtt taaaaaatgc accggcgcga gcccgagcga atttcgcgcg ggctgcgaag
     2941 aaaaagtgaa cgatgtggcg gtgaaactga gcgggtaagg ctaatggaga ttttcaacat
     3001 gggcgagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     3061 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     3121 agtgaaacgc cgtagcgccg atggtagtgt ggggtatccc catgcgagag tagggaactg
     3181 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3241 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     3301 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     3361 ttaagcagaa ggccatcctg acggatggcc ttttctgaaa ggaggaacta tatattctca
     3421 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3481 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     3541 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     3601 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3661 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3721 aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc
     3781 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3841 aaactgccgg aaatcgtcgt ggtattcact caagagcgat gaaaacgttt cagtttgctc
     3901 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3961 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     4021 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4081 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4141 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4201 agctcctgaa aatctcgata actaaaaaaa tacgcccggt agtgatctta tttcattatg
     4261 gtgaaagttg gaacctctta cgtgccgatc aacagtttta ttgttcatga ccaaaatccc
     4321 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4441 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4501 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4561 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4621 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4741 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4801 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4861 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4921 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaa
//
