LOCUS       sequence-461217-        8644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Light-inducible Cre recombinase split with REDMAP photoreceptors for
            bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8644)
  AUTHORS   Jafarbeglou F, Dunlop MJ
  TITLE     Red Light Responsive Cre Recombinase for Bacterial Optogenetics.
  JOURNAL   ACS Synth Biol. 2024 Nov 19. doi: 10.1021/acssynbio.4c00388.
  PUBMED    39558834
REFERENCE   2  (bases 1 to 8644)
  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: "ACS Synth 
            Biol. 2024 Nov 19. doi: 10.1021/acssynbio.4c00388."
COMMENT     Sequence Label: pBbS5a-OptoCre-REDMAP
FEATURES             Location/Qualifiers
     source          1..8644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(487..581)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(611..1402,1403..1471))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1234..1253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1472..1576)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1650..2732)
                     /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     2694..2713
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2733..2810)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3016..3035
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(3121..3126,3127..3144,3145..3151)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3159..3175
                     /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)."
     RBS             3992..4003
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6774..6845
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6861..6888
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7419..8369)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa
       61 acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga
      121 atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag
      181 ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta
      241 tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg
      301 cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg
      361 ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc
      421 taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctagt
      481 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
      541 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgtaaactt
      601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
      961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcggt gcctaatgag
     1621 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     1801 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     1861 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     1921 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     1981 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     2041 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     2101 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     2161 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     2221 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     2281 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     2341 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     2401 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     2461 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     2521 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     2581 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     2641 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     2701 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     2761 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     2821 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     2881 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     2941 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     3001 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     3061 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgtttagg caccccaggc
     3121 tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     3181 gaattcaaaa gatcttttaa gaaggagata tacatctggc cacctctgat gaagtcagga
     3241 agaacctgat ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc
     3301 tcctgtctgt gtgcagatcc tgggctgcct ggtgcaagct gaacgggtct ggctccggat
     3361 caagtggtgg cagccccgag gtggaggtcg acaacaacaa cgagaagccc agcgagatca
     3421 acagcttcca ccacatgatt atcagcagca gcaagaacgt gctgaagatg gaggaggtgg
     3481 aggtgtccaa gaagaggaag ttccagaccg accagagcga cgagctgagc ttattacctt
     3541 tatccaaaca cacatgcttt gccaacgtgg cttgtagcga gaacaccaac ggcaacagcg
     3601 agatcgatac cgagtacagc atgtccagct acgtgaactc caccacctcc atggagtgca
     3661 acaacgacat cgagatgaag gaagaatcca gcggcagctg cggagaggac aagatgatca
     3721 gcttcgagtc ccatttagac tatatctacg gcacccagaa tctggaggac ttctccgaga
     3781 aggtgatcga gaacattctg tacctcgacg agcaagaaga ggaggaggag gacgccaagg
     3841 gctgtagcag caacgctgcc aagttcgtgc tgtcctctgg tcgttggacc gtgaaccaag
     3901 atgatagcac tttacacgaa accaagaagc ccaccatcga ccaagaattc gagcagtact
     3961 ttagcacttt aatgctgtaa tagttaactt taaagaggag aaaggtaccg catggagaag
     4021 aagatgagcg gatctcgtcc cacacagtcc agcgagggat ctcgtagatc tcgtcactcc
     4081 gctcgtatca tcgctcagac caccgtggac gccaaactgc acgccgattt cgaggagagc
     4141 ggctcctcct ttgattactc caccagcgtg agggtgactg gtcccgtggt cgagaaccag
     4201 cctcctagga gcgacaaggt caccacaacc tacctccatc atatccagaa gggcaagctg
     4261 atccagccct ttggatgttt actggcttta gacgaaaaga ccttcaaggt catcgcctac
     4321 tccgagaacg cctccgaact gctcacaatg gcttcccacg ccgtgcctag cgtgggagag
     4381 caccccgttc tgggcatcgg caccgatatc agatctttat ttaccgctcc cagcgcttcc
     4441 gctttacaga aggctctcgg cttcggcgac gtgtctttac tgaaccctat cctcgtccat
     4501 tgtcgtacat ccgccaagcc cttctacgcc attatccata gggtgaccgg atccatcatc
     4561 atcgacttcg aacccgttaa gccctatgaa gtgcccatga cagccgccgg agctttacag
     4621 agctataagc tggccgccaa ggccatcacc agactccagt ctttacccag cggaagcatg
     4681 gagaggctgt gcgacaccat ggtgcaagaa gtcttcgagc tgactggtta cgatcgtgtc
     4741 atggcctaca agttccacga ggacgaccac ggcgaggtgg tgagcgaagt gaccaagccc
     4801 ggtttagaac cctatttagg tttacactat cccgccacag acatccccca agctgccaga
     4861 tttttattta tgaagaacaa ggtgagaatg atcgtggact gcaacgccaa gcacgctagg
     4921 gttttacaag atgagaagct gagcttcgat ttaaccctct gcggcagcac tttaagagct
     4981 ccccactctt gtcatctgca atacatggcc aatatggaca gcatcgcttc tttagtgatg
     5041 gccgtggtgg tgaatgagga ggatggagag ggcgatgctc ccgatgccac cacacagcct
     5101 cagaagagaa agaggctgtg gggtttagtg gtctgccaca acaccacccc cagatttgtc
     5161 ccttttccct tacgttatgc ttgtgaattt ttagcccaag tgtttgccat ccacgtcaac
     5221 aaggaggtcg agctggacaa ccagatggtg gagaagaaca ttttacgtac ccagacactc
     5281 ctctgcgaca tgctcatgag ggacgctccc ctcggcatcg tgagccagtc ccccaatatt
     5341 atggatttag tcaagtgcga cggcgccgcc ttattataca aggacaagat ctggaagctg
     5401 ggcaccaccc ctagcgagtt ccatttacaa gaaatcgctt cttggctgtg tgagtaccac
     5461 atggattcca ccggtttaag caccgattct ttacacgacg ctggctttcc tagggcttta
     5521 tctttaggcg acagcgtctg cggaatggct gccgttcgta tcagcagcaa agatatgatt
     5581 ttctggtttc gttcccatac cgccggcgaa gtgagatggg gcggcgccaa acacgacccc
     5641 gatgataggg acgatgccag aaggatgcac cccagatcct ccttcaaggc ttttctggag
     5701 gtggtgaaga ccagaagctt accttggaag gactacgaaa tggatgccat ccactcttta
     5761 cagctcatct tacgtaacgc ctttaaggac agcgagacca ccgacgtgaa cacaaaggtg
     5821 atctatagca agctcaacga tctcaagatt gatggaatcc aaggtaccgg aggttcagga
     5881 ggttcaaaca ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac
     5941 ctgcaagcca gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg
     6001 ctgcacagga gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg
     6061 aggagaatca gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt
     6121 gaacgcactg actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac
     6181 atcaggaacc tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt
     6241 gccagaatca gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt
     6301 ggcaggacca agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt
     6361 accaagctgg tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac
     6421 ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc
     6481 acccgggccc tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat
     6541 gactctgggc agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg
     6601 gacatggcca gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat
     6661 gtgaacattg tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg
     6721 ctgctcgagg atggggacta aggatccaaa ctcgagtaag gatctccagg catcaaataa
     6781 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     6841 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacctagg
     6901 gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc
     6961 gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag aattgccatg
     7021 attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa
     7081 gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca
     7141 ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg
     7201 ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttgaatgcac
     7261 caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata
     7321 tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg tttgttagtc
     7381 ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc
     7441 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     7501 tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     7561 gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga atctgatgta
     7621 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     7681 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     7741 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     7801 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     7861 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     7921 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     7981 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     8041 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     8101 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     8161 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     8221 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     8281 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     8341 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     8401 ctataccaat tgagatgggc tagtcaatga taattactag tccttttccc gggtgatctg
     8461 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     8521 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     8581 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     8641 ctac
//
