LOCUS       sequence-314798-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Updated light-inducible Cre recombinase split with Vivid photodimers
            for bacterial expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  AUTHORS   Sheets MB, Wong WW, Dunlop MJ
  TITLE     Light-Inducible Recombinases for Bacterial Optogenetics.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):227-235. doi: 
            10.1021/acssynbio.9b00395. Epub 2020 Jan 21.
  PUBMED    31961670
REFERENCE   2  (bases 1 to 7126)
  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"; doi: 
            "10.1021/acssynbio.9b00395"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "227-235"
COMMENT     Sequence Label: pBbS5a-Opto-Cre-Vvd-2
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(119..124,125..142,143..149)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    157..173
                     /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             852..863
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2254..2325
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2341..2368
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2899..3849)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(4611..4705)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4735..5526,5527..5595))
                     /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     5358..5377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5596..5700)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5774..6856)
                     /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     6818..6837
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6857..6934)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
       61 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgtttaggca ccccaggctt
      121 tacactttat gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaga
      181 attcaaaaga tcttttaaga aggagatata catatggcca cctctgatga agtcaggaag
      241 aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg gaagatgctc
      301 ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acgggtctgg ctccggatca
      361 agtggtggca gcggtaccca cactctctac gccccaggcg ggtacgatat tatgggctgg
      421 ctgatccaga tcatgaacag gcccaatccc caggtcgagc tgggacccgt ggatacttca
      481 tgtgcactga tactgtgcga cctgaagcag aaggatacac ctatagttta cgcttcagaa
      541 gcctttctgt acatgacagg gtattctaac gccgaggtgc tggggaggaa ctgtaggttc
      601 ctccagagtc ccgatggtat ggtgaaacct aagagtactc gcaaatatgt ggatagcaat
      661 actattaaca ccatgaggaa agccatcgac agaaacgcag aagttcaggt ggaagtggtg
      721 aactttaaga agaacggcca gcggttcgtg aactttctca caatgattcc agtgcgggac
      781 gaaaccgggg agtaccggta cagcatgggt tttcagtgcg aaaccgaacc cggataataa
      841 tagttaactt taaagaggag aaaggtaccg catgactagc aaaagacaca ctctctacgc
      901 cccaggcggg tacgatatta tgggctggct gatccagatc atgaacaggc ccaatcccca
      961 ggtcgagctg ggacccgtgg atacttcatg tgcactgata ctgtgcgacc tgaagcagaa
     1021 ggatacacct atagtttacg cttcagaagc ctttctgtac atgacagggt attctaacgc
     1081 cgaggtgctg gggaggaact gtaggttcct ccagagtccc gatggtatgg tgaaacctaa
     1141 gagtactcgc aaatatgtgg atagcaatac tattaacacc atgaggaaag ccatcgacag
     1201 aaacgcagaa gttcaggtgg aagtggtgaa ctttaagaag aacggccagc ggttcgtgaa
     1261 ctttctcaca atgattccag tgcgggacga aaccggggag taccggtaca gcatgggttt
     1321 tcagtgcgaa accgaacccg gaggtaccgg aggttcagga ggttcaaaca ggaaatggtt
     1381 ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc
     1441 tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct
     1501 gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa
     1561 tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca
     1621 agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct
     1681 gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga
     1741 catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt
     1801 gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg
     1861 gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa
     1921 gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat
     1981 ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct
     2041 ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca gggctggtgt
     2101 gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta
     2161 catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg atggggacta
     2221 aggatccaaa ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     2281 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     2341 ggctcacctt cgggtgggcc tttctgcgtt tatacctagg gtacgggttt tgctgcccgc
     2401 aaacgggctg ttctggtgtt gctagtttgt tatcagaatc gcagatccgg cttcagccgg
     2461 tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc cacgggaggc
     2521 gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc ctgtttcagg
     2581 ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat ttcatgttct
     2641 agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg ttcatctgtt
     2701 acattgtcga tctgttcatg gtgaacagct ttgaatgcac caaaaactcg taaaagctct
     2761 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     2821 atgtaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     2881 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     2941 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatacttac tttgcatgtc
     3001 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     3061 tgtttttctt agtccgttat gtaggtagga atctgatgta atggttgttg gtattttgtc
     3121 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     3181 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     3241 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     3301 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     3361 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     3421 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     3481 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     3541 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     3601 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     3661 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     3721 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     3781 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     3841 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     3901 tagtcaatga taattactag tccttttccc gggtgatctg ggtatctgta aattctgcta
     3961 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     4021 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     4081 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     4141 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     4201 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     4261 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     4321 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     4381 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     4441 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     4501 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     4561 gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca
     4621 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     4681 attactggat ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagttacca
     4741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4801 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4861 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4921 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4981 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5041 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5101 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5161 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5221 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5281 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5341 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5401 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5461 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5521 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5581 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5641 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5701 cacatttccc cgaaaagtgc cacctgacgt cggtgcctaa tgagtgagct aacttacatt
     5761 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5821 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     5881 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     5941 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     6001 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     6061 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     6121 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     6181 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     6241 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     6301 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     6361 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     6421 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     6481 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     6541 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     6601 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     6661 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     6721 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     6781 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     6841 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     6901 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     6961 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     7021 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     7081 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagccc
//
