LOCUS       Exported                5720 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Light-inducible Cre recombinase split with Vivid photodimers for 
            bacterial expression..
ACCESSION   .
VERSION     .
KEYWORDS    pBbE5a-Opto-Cre-Vvd
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  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 5720)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5720
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     22..41
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        127..157
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    165..181
                     /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             860..871
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2265..2336
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2352..2379
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3213..3307
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3337..4197)
                     /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     3960..3979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4198..4302)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4376..5458)
                     /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     5420..5439
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5459..5536)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
       61 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg tttaggcacc
      121 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga gcggataaca
      181 atttcagaat tcaaaagatc ttttaagaag gagatataca tatggccacc tctgatgaag
      241 tcaggaagaa cctgatggac atgttcaggg acaggcaggc cttctctgaa cacacctgga
      301 agatgctcct gtctgtgtgc agatcctggg ctgcctggtg caagctgaac gggtctggct
      361 ccggatcaag tggtggcagc ggtacccaca ctctctacgc cccaggcggg tacgatatta
      421 tgggctggct gatccagatc atgaacaggc ccaatcccca ggtcgagctg ggacccgtgg
      481 atacttcatg tgcactgata ctgtgcgacc tgaagcagaa ggatacacct atagtttacg
      541 cttcagaagc ctttctgtac atgacagggt attctaacgc cgaggtgctg gggaggaact
      601 gtaggttcct ccagagtccc gatggtatgg tgaaacctaa gagtactcgc aaatatgtgg
      661 atagcaatac tattaacacc atgaggaaag ccatcgacag aaacgcagaa gttcaggtgg
      721 aagtggtgaa ctttaagaag aacggccagc ggttcgtgaa ctttctcaca atgattccag
      781 tgcgggacga aaccggggag taccggtaca gcatgggttt tcagtgcgaa accgaacccg
      841 gataataata gttaacttta aagaggagaa aggtaccgca tgactagcaa aagacacact
      901 ctctacgccc caggcgggta cgatattatg ggctggctga tccagatcat gaacaggccc
      961 aatccccagg tcgagctggg acccgtggat acttcatgtg cactgatact gtgcgacctg
     1021 aagcagaagg atacacctat agtttacgct tcagaagcct ttctgtacat gacagggtat
     1081 tctaacgccg aggtgctggg gaggaactgt aggttcctcc agagtcccga tggtatggtg
     1141 aaacctaaga gtactcgcaa atatgtggat agcaatacta ttaacaccat gaggaaagcc
     1201 atcgacagaa acgcagaagt tcaggtggaa gtggtgaact ttaagaagaa cggccagcgg
     1261 ttcgtgaact ttctcacaat gattccagtg cgggacgaaa ccggggagta ccggtacagc
     1321 atgggttttc agtgcgaaac cgaacccgga taaggtaccg gaggttcagg aggttcaaac
     1381 aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc
     1441 agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg
     1501 agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc
     1561 agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact
     1621 gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac
     1681 ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc
     1741 agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc
     1801 aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg
     1861 gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc
     1921 cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc
     1981 ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg
     2041 cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccag ggacatggcc
     2101 agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt
     2161 gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgag
     2221 gatggggact aaggatccaa actcgagtaa ggatctccag gcatcaaata aaacgaaagg
     2281 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     2341 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg actagtgctt ggattctcac caataaaaaa cgcccggcgg
     3241 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     3301 aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4321 gccacctgac gtcggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4441 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4501 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4561 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     4621 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     4681 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     4741 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     4801 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     4861 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     4921 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     4981 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     5041 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     5101 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     5161 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5221 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5281 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5341 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5401 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5461 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5521 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     5581 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     5641 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5701 gcgctcatga gcccgaagtg
//
