LOCUS       sequence-372297-        7285 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Opto-T7RNAP*(563-F2), half expr..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7285)
  AUTHORS   Baumschlager A, Aoki SK, Khammash M
  TITLE     Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for 
            Precise Spatiotemporal Gene Expression Control.
  JOURNAL   ACS Synth Biol. 2017 Nov 17;6(11):2157-2167. doi: 
            10.1021/acssynbio.7b00169. Epub 2017 Oct 18.
  PUBMED    29045151
REFERENCE   2  (bases 1 to 7285)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00169"; journalName: "ACS Synth Biol"; date: 
            "2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2157-2167"
COMMENT     Sequence Label: pAB166
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      64..286
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(778..872)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(893..1552)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1467..1486
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1553..1655)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3491..3940
                     /codon_start=1
                     /product="""negative Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=nMagHigh1
                     /note="rapidly heterodimerizes with pMag or pMagFast2 upon 
                     blue light irradiation"
                     /translation="HTLYAPGGYDIMGYLDQIGNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTIRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTIIPVRDETGEYRYSMGFQCETE"
     terminator      3956..4042
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             4645..5094
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMagFast2
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with fast switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALVLCDLKQK
                     DTPVVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     terminator      6084..6170
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(6351..7285,1..16))
                     /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 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
       61 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
      121 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
      181 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
      241 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
      301 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
      361 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
      421 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
      481 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
      541 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
      601 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
      661 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
      721 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
      781 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
      841 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc
      901 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
      961 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     1021 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     1081 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     1141 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     1201 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     1261 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     1321 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     1381 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     1441 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     1501 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     1561 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     1621 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc cagatatcga
     1681 cgtctagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     1741 tctactgttt ctccataccc gtttttttgg aaggaggtaa agatctatga acacgattaa
     1801 catcgctaag aacgacttct ctgacatcga actggctgct atcccgttca acactctggc
     1861 tgaccattac ggtgagcgtt tagctcgcga acagttggcc cttgagcatg agtcttacga
     1921 gatgggtgaa gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc
     1981 ggataacgct gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat
     2041 caacgactgg tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt
     2101 cctgcaagaa atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg
     2161 cctaaccagt gctgacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat
     2221 tgaggacgag gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa
     2281 cgttgaggaa caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt
     2341 tgtcgaggct gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca
     2401 taaggaagac tctattcatg taggagtacg ctgcatcgag atgctcattg agtcaaccgg
     2461 aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga
     2521 actcgcacct gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc
     2581 tccgatgttc caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg
     2641 ctattgggct aacggtcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact
     2701 gatgcgctac gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa
     2761 caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa
     2821 gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga
     2881 agacatcgac atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta
     2941 ccgcaaggac aaggctcgca agtctcgccg tatcagcctt gagttcatgc ttgagcaagc
     3001 caataagttt gctaaccata aggccatctg gttcccttac aacatggact ggcgcggtcg
     3061 tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gactgcttac
     3121 gctggcgaaa ggtaaaccaa tcggtaagga aggttactac tggctgaaaa tccacggtgc
     3181 aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa
     3241 ccacgagaac atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca
     3301 agattctccg ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg
     3361 cctgagctat aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca
     3421 cttctccgcg atgctccgag atgaggtagg tggtcgcgcg gttaacttgc ttcctggcgg
     3481 ttctggaggt cacactcttt acgcccctgg aggatacgac attatgggat atttggatca
     3541 gattgggaac cgcccaaacc ctcaggtcga actggggcct gtggacacgt catgtgccct
     3601 gatcctgtgc gatctgaagc aaaaggacac tccgatcgtc tacgcctcgg aagccttctt
     3661 gtatatgacc ggatacagca atgcagaggt gctcggcagg aactgcagat tcctgcagtc
     3721 ccccgacggg atggtgaaac caaagtcgac tcgcaaatat gtggactcga acacgatcaa
     3781 caccatccgg aaggccatcg accggaacgc cgaggtccag gtggaggtgg tcaactttaa
     3841 gaagaacggc cagcggttcg tgaactttct gaccatcatt ccggtccggg atgaaaccgg
     3901 agagtacaga tactccatgg gattccagtg cgaaaccgaa taacctagga ggcatcaaat
     3961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4021 cgctctcctg agtaggacaa atccgccgcc ctagacctag ctgcagctag ggtacgggtt
     4081 ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg
     4141 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc
     4201 ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg
     4261 cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa
     4321 tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct
     4381 gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     4441 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat gcgccgatca
     4501 acgtctcatt ttcgccagat atcggcgcgc cgcttcgtag catttttatc cataagatta
     4561 gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt
     4621 tggaaggagg taaattaatt aatgcacact ctttacgccc ctggaggata cgacattatg
     4681 ggatatttgc ggcagattag gaaccgccca aaccctcagg tcgaactggg gcctgtggac
     4741 acgtcatgtg ccctggtcct gtgcgatctg aagcaaaagg acactccggt ggtctacgcc
     4801 tcggaagcct tcttgtatat gaccggatac agcaatgcag aggtgctcgg caggaactgc
     4861 agattcctgc agtcccccga cgggatggtg aaaccaaagt cgactcgcaa atatgtggac
     4921 tcgaacacga tcaacaccat gcggaaggcc atcgaccgga acgccgaggt ccaggtggag
     4981 gtggtcaact ttaagaagaa cggccagcgg ttcgtgaact ttctgaccat gattccggtc
     5041 cgggatgaaa ccggagagta cagatactcc atgggattcc agtgcgaaac cgaaggcggt
     5101 tctggaggta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag
     5161 attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag
     5221 aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg
     5281 ctggcttacg gtgttactcg cagtgtgact aagagttcag tcatgacgct ggcttacggg
     5341 tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat
     5401 tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg
     5461 atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag
     5521 tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc
     5581 aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag
     5641 aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc
     5701 attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct
     5761 aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag
     5821 aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac
     5881 gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat
     5941 gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa
     6001 atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc
     6061 gcgttcgcgt aatctagagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     6121 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     6181 agacctagct gcagctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg
     6241 ctaggatcca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     6301 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     6361 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     6421 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     6481 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     6541 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     6601 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     6661 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     6721 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     6781 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     6841 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     6901 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     6961 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     7021 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     7081 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     7141 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     7201 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     7261 atcgctagtt catttgcttt gaaaa
//
