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     14..33
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        119..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      2257..2328
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2344..2371
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2529..3117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2609..2628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3205..3299
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3329..4189)
                     /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     3952..3971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4190..4294)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4368..5450)
                     /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     5412..5431
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5451..5528)
                     /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 gataaggtac cggaggttca ggaggttcaa acaggaaatg
     1381 gttccctgct gaacctgagg atgtgaggga ctacctcctg tacctgcaag ccagaggcct
     1441 ggctgtgaag accatccaac agcacctggg ccagctcaac atgctgcaca ggagatctgg
     1501 cctgcctcgc ccttctgact ccaatgctgt gtccctggtg atgaggagaa tcagaaagga
     1561 gaatgtggat gctggggaga gagccaagca ggccctggcc tttgaacgca ctgactttga
     1621 ccaagtcaga tccctgatgg agaactctga cagatgccag gacatcagga acctggcctt
     1681 cctgggcatt gcctacaaca ccctgctgcg cattgccgaa attgccagaa tcagagtgaa
     1741 ggacatctcc cgcaccgatg gtgggagaat gctgatccac attggcagga ccaagaccct
     1801 ggtgtccaca gctggtgtgg agaaggccct gtccctgggg gttaccaagc tggtggagag
     1861 atggatctct gtgtctggtg tggctgatga ccccaacaac tacctgttct gccgggtcag
     1921 aaagaatggt gtggctgccc cttctgccac ctcccaactg tccacccggg ccctggaagg
     1981 gatctttgag gccacccacc gcctgatcta tggtgccaag gatgactctg ggcagagata
     2041 cctggcctgg tctggccact ctgccagagt gggtgctgcc agggacatgg ccagggctgg
     2101 tgtgtccatc cctgaaatca tgcaggctgg tggctggacc aatgtgaaca ttgtgatgaa
     2161 ctacatcaga aacctggact ctgagactgg ggccatggtg aggctgctcg aggatgggga
     2221 ctaaggatcc aaactcgagt aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg
     2281 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     2341 actggctcac cttcgggtgg gcctttctgc gtttatacct agggcgttcg gctgcggcga
     2401 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2461 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2521 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2581 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2641 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2701 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2761 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2821 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2881 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2941 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3001 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3061 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3121 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3181 attttggtca tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     3241 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     3301 aagcgagctc gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4321 acgtcggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4381 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4441 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4501 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4561 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4621 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta
     4681 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4741 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4801 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4861 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4921 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4981 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5041 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5101 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5161 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5221 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5281 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5341 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5401 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5461 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5521 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5581 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5641 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5701 gagcccgaag tggcgagccc
//
