LOCUS       pDest_PANCEd_Ara        4969 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  AUTHORS   Nicholas Southern
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="recombinant plasmid"
     primer_bind     154..171
                     /label=L4440 vector, forward primer
                     /note="L4440 vector, forward primer"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(379..1461)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1423..1442
                     /label=LacI, reverse primer
                     /note="LacI, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(1462..1539)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        1584..1613
                     /note="strong E. coli promoter; hybrid between the trp
                     andlac UV5 promoters"
                     /note="color: #ffffff"
     protein_bind    1621..1633
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     terminator      1638..1641
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
                     /note="color: #ffffff"
     terminator      1638..1639
                     /note="color: #ffffff"
     misc_feature    1645..1658
                     /label=sd2 RBS
                     /note="color: #a6acb3"
     misc_feature    1659..2978
                     /label=AraC
                     /note="color: #993366"
     variation       1668..1670
                     /label=E3K
                     /note="color: #a6acb3"
     misc_feature    1668
                     /label=G to A
                     /note="color: #a6acb3"
     variation       1710..1712
                     /label=Silent Mutation
                     /note="color: #a6acb3"
     misc_feature    1712
                     /label=G to A
                     /note="color: #a6acb3"
     variation       1926..1928
                     /label=Silent Mutation
                     /note="color: #a6acb3"
     misc_feature    1928
                     /label=C to A
                     /note="color: #a6acb3"
     variation       2097..2099
                     /label=R146L
                     /note="color: #c0c0c0"
     misc_feature    2098
                     /label=G to T
                     /note="color: #a6acb3"
     variation       2115..2117
                     /label=A152T
                     /note="color: #a6acb3"
     misc_feature    2115
                     /label=G to A
                     /note="color: #a6acb3"
     misc_feature    2117
                     /label=G to A
                     /note="color: #a6acb3"
     motif           2172..2177
                     /label=linker
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     CDS             2178..2600
                     /label=LOV2_ecoli
                     /note="/created_by=User"
                     /note="color: #993366; direction: RIGHT"
     motif           2601..2606
                     /label=linker
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     variation       2718..2720
                     /label=Silent Mutation
                     /note="color: #c0c0c0"
     misc_feature    2720
                     /label=G to T
                     /note="color: #a6acb3"
     variation       2847..2849
                     /label=R251L
                     /note="color: #c0c0c0"
     misc_feature    2848
                     /label=G to T
                     /note="color: #a6acb3"
     misc_feature    2979..2981
                     /standard_name="pr"
                     /label=pr
                     /note="color: #a6acb3"
     misc_feature    2986..3001
                     /standard_name="retained"
                     /label=retained
                     /note="color: #a6acb3"
     misc_feature    3002..3004
                     /standard_name="J23107 promoter"
                     /label=J23107 promoter
                     /note="color: #a6acb3"
     primer_bind     complement(3019..3036)
                     /note="For vectors with E. coli araBAD promoter,
                     reverseprimer"
                     /note="color: #a020f0; direction: LEFT"
     terminator      3189..3275
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="transcription terminator T1 from the E. coli
                     rrnBgene"
                     /note="color: #ffffff"
     terminator      3367..3394
                     /note="transcription terminator T2 from the E. coli
                     rrnBgene"
                     /note="color: #ffffff"
     terminator      complement(3414..3508)
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff; direction: LEFT"
     terminator      3414..3508
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
     primer_bind     3414..3445
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(3491..3519)
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(3529..4188)
                     /codon_start=1
                     /note="color: #993366"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3529..4188)
                     /gene="CAT/CamR"
                     /label=CAT/CamR
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(3752..3757)
                     /codon_start=1
                     /note="color: #993366"
                     /translation="MF"
     misc_feature    3752..3757
                     /label=Geneious type: Concatenated sequence
                     /label=nonstandard type: Concatenated sequence
                     /note="color: #a6acb3"
     marker          complement(3752..3757)
                     /direction=LEFT
                     /label=Geneious type: marker
                     /note="color: #a6acb3"
     gene            complement(3752..3757)
                     /gene="CAT/CamR"
                     /label=CAT/CamR
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     4103..4122
                     /label=Chloramphenicol resistance gene, reverse primer
                     /note="Chloramphenicol resistance gene, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(4189..4291)
                     /note="promoter of the E. coli cat gene
                     encodingchloramphenicol acetyltransferase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     4292..4330
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     4870..4889
                     /label=pBR322 origin, forward primer
                     /note="pBR322 origin, forward primer"
                     /note="color: #a020f0; direction: RIGHT"
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 ccagagcgat 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 actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4261 gtgaaagttg gaacctctta cgtgccgatc aacagtttta ttgttcatga ccaaaatccc
     4321 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 aagggatcct 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
//
