LOCUS       sequence-339482-        6031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Renilla luciferase reporter plasmid; Giles integrating..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  AUTHORS   Bosch B, DeJesus MA, Poulton NC, Zhang W, Engelhart CA, Zaveri A, 
            Lavalette S, Ruecker N, Trujillo C, Wallach JB, Li S, Ehrt S, Chait 
            BT, Schnappinger D, Rock JM
  TITLE     Genome-wide gene expression tuning reveals diverse vulnerabilities 
            of M. tuberculosis.
  JOURNAL   Cell. 2021 Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033.
  PUBMED    34297925
REFERENCE   2  (bases 1 to 6031)
  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"; journalName: "Cell. 2021 
            Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033."
COMMENT     Sequence Label: plRL118
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      92..119
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(165..170,171..188,189..195)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    201..220
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(1182..1201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1183..1201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1220..1236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1220..1236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1233..1255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1244..1260
                     /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)."
     promoter        complement(join(1268..1274,1275..1292,1293..1298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1313..1334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1451..1468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1622..2210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(4227..4331)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4358..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4445..4464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4655..4676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4939..4961
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4953..4970
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4954..4970
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(5167..5253)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5298..5867)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     terminator      complement(join(5989..6031,1..4))
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tttgatgcct ggcagtcgat cgtacgctag ttaactacgt cgacatcgat caatatgact
       61 agtagatcct ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
      121 gcgtttctac aaactcttcc tgtcgtcata tctagacccc aggcttgaca ctttatgctt
      181 ccggctcgta taatgtgtgg aattgtgagc gctcacaatt cggatccagc aggaggtatc
      241 tccatggcga gcaaggtcta cgaccccgag caacgtaagc gcatgatcac cggcccccaa
      301 tggtgggcgc ggtgcaagca gatgaatgtg ctggactcgt tcatcaatta ctatgactcc
      361 gaaaaacacg ccgaaaacgc cgtgattttc ctgcacggca atgccgcctc gtcgtacctc
      421 tggcggcacg tggtgcccca catcgagccc gtggcccggt gcatcatccc ggacctgatc
      481 ggcatgggca agtcgggcaa gtcgggtaac ggcagctacc gcttgctgga ccactataaa
      541 tacttgaccg cctggtttga attgctgaac ttgccgaaaa agatcatttt cgtgggccat
      601 gactggggtg cctgcctggc ctttcactat agctacgagc accaagacaa gattaaggcg
      661 atcgtccacg cggagagcgt ggtggatgtg atcgagagct gggacgagtg gccggacatc
      721 gaggaggaca ttgccctgat caaaagcgaa gaaggcgaga agatggtcct ggagaacaac
      781 ttcttcgtgg agaccatgct gccatcgaag atcatgcgca aactcgaacc ggaggagttc
      841 gccgcgtacc tcgaaccctt taaggagaaa ggggaggtgc gccggccgac cttgtcgtgg
      901 ccccgcgaaa tcccgctggt caagggcggc aagccggacg tcgtccagat cgtgcggaac
      961 tacaacgcgt atctccgggc gtcggacgac ctgcccaaga tgttcattga aagcgatcca
     1021 ggatttttct cgaacgccat cgtggagggt gcgaagaagt tcccgaacac cgagttcgtc
     1081 aaagtcaagg gactgcactt cagccaggag gacgcccccg acgagatggg gaaatatatc
     1141 aagtcgttcg tggagcgcgt cttgaagaac gaacagtagt tccctatagt gagtcgtatt
     1201 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1261 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1321 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1441 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2281 tcatgagatt atcaaaaagg atcttcacct agatcctttt ggaatgatga tcgccgcggt
     2341 gacacaatcg gcgtacgcgg acgcccggac gtgaccggag gcgaccgtac acctatcccg
     2401 accgggcact attcggcgtg gatccccacg cacccggcga cccccgagag gacaccgcaa
     2461 tggggcgacg cggcaaccgc gaaggcacat accggcaccg gccggacggg cgctgggagg
     2521 cccggttgct ctatgacgac ccggtcaccg gtgaacgccg ccgggcatcg ttctacggca
     2581 agacgaagac ggcggtacgg gccaaaatgc gcgaagctat cgaccgcctc gacgccgacg
     2641 ccccggtgcg tgacgccgcg gtgaccgtgg ccgattggat gcgccggtgg tgcaccacca
     2701 ccctgttggt ctccggccgc gccgacagca ccaaatcgtt gtacggcaca ctgtcgcgcc
     2761 aatacgtcga acccgccccg ttcggcgcga tccggttgga ccgcctgcgc acatccgatg
     2821 tcgaggcgtt ggtgctgcgc cttcaacagg ccacccgccc gggccgcgac gggacgccgg
     2881 tgcgcgcggt cgccgacagc acgatccggt cgacctacac cgtgttgcgc caggcgctcg
     2941 acggcgcggt gcgcgacggc ctgatggcgc gcaaccctgc ggcggccatg aagcggcccg
     3001 ggttgtcgcg cccggaagtc cggcggatca acgacgcgca gttggccgac ctgttggccg
     3061 ccaccgccgg atcccgctac caccgcgccc tgttgctgat cgcggcgacc ggactgcgcg
     3121 ccggggaggc gttggccctt cggtgggacg ccgtcgacct cgacgccggg tcgctgcgcg
     3181 tggtggccac cgtcggccgc gtcgagcgtc gcctgcgcct cacgacgccg aagacggaac
     3241 gatcccgccg gacggtgccg ttgacgccgg acatggtgtc gatgctgcgt cggcaccgcg
     3301 ccgaccagtt agctgaacgc ctgcgcgcgg ccaacgtgtg ggaggaccac gggttggtgt
     3361 tcccgtcggc gttcggccgc ccggtggatc cgcgcaacct gttgcgggtg gtggagcagg
     3421 ccgccgcgca ggttggcctc gagggcgtcg gcgtgcacac cctgcgtcac gccgcggcgg
     3481 tggcgatgct cgaggcgggc atccacatcc gcgcggtcgc cgacatcctc gggcattcgt
     3541 cggtggcggt cacgggggac atctacggcc acacgtccga cgcgaccgcg cgggccgccg
     3601 tcgacgtgtt gtcggcgcgc ctgcggatcg cggacggcag cggacagtct tagcgagctc
     3661 gacggacgcg gacggacgcc ggagcaattg gggtcaaaat tggggtcaaa cagaaaaagg
     3721 ggcggcttcc cgtctccgga aaaccgcccc tgatctgcgt cgggctgaca ggatttgaac
     3781 ctgcgacccc ttgaccccca gtcattcaaa aggcatccgc gtggatccgc ccgggtaccg
     3841 aatccgcagg tcacgcccgg ttgcgcacaa tggcggacat cggcgaatcg ccccgaaccc
     3901 cgatgcaatt ggggtcaaaa ttggggtcaa accggcgggg gtgaccgctt aactctgcga
     3961 atacgtgtag gtcggcgtca cgtcgacctg gcccggcccc gcggagttga tcgtcaaaaa
     4021 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4081 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4141 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4201 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4261 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4321 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     4381 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4441 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4501 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4561 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4621 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4681 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4741 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4801 acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4861 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4921 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     4981 tcggtacccg gggatccgat atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5101 gttgcctgac tccccgacgt ggccgaccag cccgtcctgc gcaacgttca aatccgctcc
     5161 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     5221 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
     5281 ggggagagat gcatctatca ggggcagggc atgctcatgt agagcgcctg ctcgccgtcc
     5341 gaggcggtgc cgtcgtacag ggcggtgtcc aggccgcaga gggtgaaccc catccgccgg
     5401 tacgcgtgga tcgccggtgc gttgacgttg gtgacctcca gccagaggtg cccggcgccc
     5461 cgctcgcggg cgaactccgt cgcgagcccc atcaacgcgc gcccgacccc gtgcccccgg
     5521 tgctccgggg cgacctcgat gtcctcgacg gtcagccggc ggttccagcc ggagtacgag
     5581 acgaccacga agcccgccag gtcgccgtcg tccccgtacg cgacgaacgt ccgggagtcc
     5641 gggtcgccgt cctccccgtc gtccgattcg tcgtccgatt cgtcgtcggg gaacaccttg
     5701 gtcaggggcg ggtccaccgg cacctcccgc agggtgaagc cgtccccggt ggcggtgacg
     5761 cggaagacgg tgtcggtggt gaaggaccca tccagtgcct cgatggcctc ggcgtccccc
     5821 gggacactgg tgcggtaccg gtaagcagtg tcgtcaagag tggtcatagg tatatctcct
     5881 tcgcatgcct cccagattat atcactcgag ctagcggtgt caaggcgcgc caggacaccg
     5941 cccccggcgc ctgacgcccc tctagctgat caccgcggcc atgatggcat aaaacgaaag
     6001 gcccagtctt tcgactgagc ctttcgtttt a
//
