LOCUS       sequence-372120-        7285 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Opto-T7RNAP*(69), 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: pAB170
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             2009..2458
                     /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      2474..2560
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3163..3612
                     /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=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     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 gccggcggtt ctggaggtca cactctttac gcccctggag gatacgacat
     2041 tatgggatat ttggatcaga ttgggaaccg cccaaaccct caggtcgaac tggggcctgt
     2101 ggacacgtca tgtgccctga tcctgtgcga tctgaagcaa aaggacactc cgatcgtcta
     2161 cgcctcggaa gccttcttgt atatgaccgg atacagcaat gcagaggtgc tcggcaggaa
     2221 ctgcagattc ctgcagtccc ccgacgggat ggtgaaacca aagtcgactc gcaaatatgt
     2281 ggactcgaac acgatcaaca ccatccggaa ggccatcgac cggaacgccg aggtccaggt
     2341 ggaggtggtc aactttaaga agaacggcca gcggttcgtg aactttctga ccatcattcc
     2401 ggtccgggat gaaaccggag agtacagata ctccatggga ttccagtgcg aaaccgaata
     2461 acctaggagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2521 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacctagct
     2581 gcagctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt
     2641 atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga
     2701 attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg
     2761 attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa
     2821 gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc
     2881 tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt
     2941 tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat
     3001 ctgtgcatgc gccgatcaac gtctcatttt cgccagatat cggcgcgccg cttcgtagca
     3061 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
     3121 tctccatacc cgtttttttg gaaggaggta aattaattaa tgcacactct ttacgcccct
     3181 ggaggatacg acattatggg atatttgcgg cagattagga accgcccaaa ccctcaggtc
     3241 gaactggggc ctgtggacac gtcatgtgcc ctgatcctgt gcgatctgaa gcaaaaggac
     3301 actccgatcg tctacgcctc ggaagccttc ttgtatatga ccggatacag caatgcagag
     3361 gtgctcggca ggaactgcag attcctgcag tcccccgacg ggatggtgaa accaaagtcg
     3421 actcgcaaat atgtggactc gaacacgatc aacaccatgc ggaaggccat cgaccggaac
     3481 gccgaggtcc aggtggaggt ggtcaacttt aagaagaacg gccagcggtt cgtgaacttt
     3541 ctgaccatga ttccggtccg ggatgaaacc ggagagtaca gatactccat gggattccag
     3601 tgcgaaaccg aaggcggttc tggaggtgcc aagcctctca tcactaccct actccctaag
     3661 atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg
     3721 acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag
     3781 accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca
     3841 atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag
     3901 cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa
     3961 gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg
     4021 tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc
     4081 attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac
     4141 tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg
     4201 ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc
     4261 attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac
     4321 agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt
     4381 aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta
     4441 atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc
     4501 ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct
     4561 gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc
     4621 atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg
     4681 gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc
     4741 aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg
     4801 aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag
     4861 ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact
     4921 tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg
     4981 gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc
     5041 tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac
     5101 ttgcttccta 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
//
