LOCUS       sequence-486033-        4966 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Expression Plasmid containing a LacI repressed sd2 AraC R2-LOV 
            POGO-PANCE/RAMPhaGE Linker Library variant cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4966)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 4966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDest_PANCEd_AraC_R2_LDelK
FEATURES             Location/Qualifiers
     source          1..4966
                     /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(3016..3033)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3186..3272
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3364..3391
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3411..3505)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3526..4185)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LLSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4100..4119
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4186..4288)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4378..4966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4867..4886
                     /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 caaattagcc acaacgctgg aacgcattga aaagaatttc gtaatcacag
     2221 acccgcgcct tcccgacaat ccaattattt ttgcgtccga tagcttcctg caattaaccg
     2281 aatacagccg cgaagaaatt ctgggtcgta attgtcgctt ccttcagggg ccagagactg
     2341 accgtgctac ggtacgcaaa atccgcgacg caatcgacaa tcaaacggaa gtcacggttc
     2401 agttgattaa ctatacgaag agcggaaaaa aattctggaa tttatttcac ttgcagccta
     2461 tgcgtgacca gaagggcgat gtccagtatt tcattggcgt tcagcttgat ggtaccgagc
     2521 atgttcgcga tgctgcggag cgtgaaggtg taatgttaat taaaaagact gctgaaaaca
     2581 ttgatgaggc ggccaaaggg agcctgcatc cgccgatgga taaccgcgtg cgcgaagcgt
     2641 gccagtatat tagcgatcat ctggcggata gcaactttga tattgcgagc gtggcgcagc
     2701 atgtgtgcct gagccctagc cgcctgagcc atctgtttcg ccagcagctg ggcattagcg
     2761 tgctgagctg gcgcgaagat cagcgcatta gccaggcgaa actgctgctg agcaccaccc
     2821 gcatgccgat tgcgaccgtg ggcctcaacg tgggctttga tgatcagctg tattttagcc
     2881 gcgtgtttaa aaaatgcacc ggcgcgagcc cgagcgaatt tcgcgcgggc tgcgaagaaa
     2941 aagtgaacga tgtggcggtg aaactgagcg ggtaaggcta atggagattt tcaacatggg
     3001 cgagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     3061 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     3121 gaaacgccgt agcgccgatg gtagtgtggg gtatccccat gcgagagtag ggaactgcca
     3181 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     3241 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     3301 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     3361 agcagaaggc catcctgacg gatggccttt tctgaaagga ggaactatat attctcacca
     3421 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3481 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     3541 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     3601 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     3661 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     3721 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
     3781 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     3841 ctgccggaaa tcgtcgtggt attcactcaa gagcgatgaa aacgtttcag tttgctcatg
     3901 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     3961 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     4021 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     4081 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     4141 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     4201 tcctgaaaat ctcgataact aaaaaaatac gcccggtagt gatcttattt cattatggtg
     4261 aaagttggaa cctcttacgt gccgatcaac agttttattg ttcatgacca aaatccctta
     4321 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4381 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4441 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4501 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4561 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4621 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4681 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4741 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4801 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4861 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4921 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaa
//
