LOCUS       sequence-371957-        5841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LAA-tagged mCherry under T7 promoter control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5841)
  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 5841)
  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.7b00169"; journalName: "ACS Synth Biol"; date: 
            "2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2157-2167"
COMMENT     Sequence Label: pAB143
FEATURES             Location/Qualifiers
     source          1..5841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(23..100)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     306..325
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1211..1255
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1294..1312)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1308..1355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1399..1854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1696..1717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1969..2760,2761..2829))
                     /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     2592..2611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2830..2922)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3003..3591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3492..3511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3745..3762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3777..3916
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3842..3861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4002..4024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4018..4209)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4781..5841,1..22))
                     /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     join(5825..5841,1..3)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
       61 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
      121 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
      181 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
      241 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
      301 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
      361 gccacgatgc gtccggcgta gcctaggtaa tacgactcac tatagggaga ggatcctgtc
      421 aatttccgcg atagaggagg taaagcatat ggtttcaaaa ggcgaagaag acaacatggc
      481 gattatcaag gaatttatgc gtttcaaggt ccacatggaa ggcagcgtca atggtcacga
      541 atttgaaatt gaaggcgaag gtgaaggccg tccgtatgaa ggcacccaga cggcaaaact
      601 gaaggtcacc aaaggcggtc cgctgccgtt tgcttgggat attctgtcac cgcaattcat
      661 gtatggttcg aaagcgtacg ttaagcatcc ggccgatatc ccggactatc tgaaactgtc
      721 ctttccggaa ggcttcaaat gggaacgtgt tatgaacttc gaagatggcg gtgtggttac
      781 cgtcacgcag gatagctctc tgcaagacgg tgaatttatt tataaagtga agctgcgcgg
      841 caccaatttc ccgagcgatg gtccggttat gcagaaaaag acgatgggct gggaagcgag
      901 ttccgaacgt atgtacccgg aagacggtgc cctgaaaggc gaaatcaagc agcgcctgaa
      961 actgaaggat ggcggtcact atgacgcaga agtgaaaacc acgtacaagg ctaaaaagcc
     1021 ggtccaactg ccgggtgcat acaacgtgaa catcaagctg gatatcacca gccataacga
     1081 agactatacg atcgttgaac agtacgaacg tgcagaaggc cgccactcta ccggcggtat
     1141 ggatgaactg tacaaagccg cgaatgacga gaattacgct ctggccgctt aaggtaccct
     1201 cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgtctac
     1261 tagtcgcagc ttaattaacc taaactgctg ccaccgctga gcaataacta gcataacccc
     1321 ttggggcctc taaacgggtc ttgaggggtt ttttgctagc gaaaggagga gtcgactata
     1381 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttctggcg
     1861 gcacgatggc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2821 aatactcata ctcttccttt ttcaatcatg attgaagcat ttatcagggt tattgtctca
     2881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggtcat gaccaaaatc
     2941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3121 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3781 atgcggtatt ttctccttac acatctgtgc ggtatttcac accgcaatgg tgcactctca
     3841 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3901 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3961 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4021 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     4081 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     4141 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     4201 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     4261 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     4321 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     4381 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     4441 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     4501 ttacgaaaca cggaaaccga agaccattca tattgttgct caggtcgcag acgttttgca
     4561 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4621 accccgccag cctagccggg tcctcaacga caggagcacg atcatgctag tcatgccccg
     4681 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     4741 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4861 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     4921 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     4981 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     5041 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     5101 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     5161 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     5221 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     5281 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     5341 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     5401 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     5461 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     5521 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     5581 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     5641 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     5701 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     5761 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     5821 caccggcata ctctgcgaca t
//
