LOCUS       sequence-97759-e        5655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PI driving expression FruR-L, and PI driving expression of 
            RbsR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5655)
  AUTHORS   Shis DL, Hussain F, Meinhardt S, Swint-Kruse L, Bennett MR
  TITLE     Modular, Multi-Input Transcriptional Logic Gating with Orthogonal 
            LacI/GalR Family Chimeras.
  JOURNAL   ACS Synth Biol. 2014 Sep 19;3(9):645-51. doi: 10.1021/sb500262f. 
            Epub 2014 Jul 28.
  PUBMED    25035932
REFERENCE   2  (bases 1 to 5655)
  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/sb500262f";
            journalName: "ACS Synth Biol"; date: "2014-09-19- 19"; volume: "3"; 
            issue: "9"; pages: "645-51"
COMMENT     Sequence Label: pZM3[I:FruR-L,I:RbsR-L]
FEATURES             Location/Qualifiers
     source          1..5655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1169..1246)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1290..1308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1373..1450
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2959..3150
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3144..3166)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3252..3394
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3307..3326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3409..3426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3580..4168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3660..3679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4352..5011)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4926..4945
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(5189..5644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5326..5347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5538..5557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttagcta cggctgagca
      181 cgccgcggcg atagagatta cgtttaatgc gcgttaaacc aggttttggc ttacgcggtt
      241 cgtccaggct tgccaggaca atctccagca cacgctctgc gacatcgcgg tgacgttgag
      301 ccactgccag caccggacac tgtaagaagt cgagcagttc gttatcgcca aaggtggcaa
      361 ttgccaggtc agaaggcagt ttgccgtcgc gacgcagcgt gacatccatc actccttgca
      421 acaacgcaaa cgacgttgtg aacagcgcct gcggcatcgg atgcgtttcc agccattttt
      481 cgaataactg ggcagccgcc tcccgctcat agctgttggc atacaggaaa tgcacttcgc
      541 gcggatcatc tttccaggca gtacggaaac cttgttcacg caggaagctg acagaaagct
      601 ccggtagcgc accaagataa agcaccgtct cggcgggaaa cttacgtaac tcttccgcca
      661 gcatttcggc atcatcctga tcggcaccaa ccacgctggt gaagtgttca cgatcgaggg
      721 cgcggtccag cgcgacaatc gggaacgggt cgttagccca gcgttgataa aaaggatgct
      781 caggaggcaa cgacgtcgaa acaataatgg catcaacctg acgctgtaaa aggtgctcaa
      841 tgcaccgcat ttcgttgtct ggctgatctt ctgagcaggc aatcagcagt tgataacccc
      901 gttgccgcgc ctggcgttca agatagttag cgatgcgggt atagctggtg ttctccagat
      961 cggggatcac aagaccaata gaacggctac ttttgcccgc cagttgttgt gccacgcggt
     1021 tgggaatgta attgagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1081 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     1141 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     1201 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtcgcta gcctaagaaa
     1261 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1321 acctcgacaa ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttggta ccgacaccat
     1381 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     1441 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1501 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1561 agtggaagcg gcgatggcgg agctcaatta cattcccaac cgcgtggcac aacaactggc
     1621 gggcaaagcg agccatacca ttggcatgtt gatcactgcc agtaccaatc ctttctattc
     1681 agaactggtg cgtggcgttg aacgcagctg cttcgaacgc ggttatagtc tcgtcctttg
     1741 caataccgaa ggcgatgaac agcggatgaa tcgcaatctg gaaacgctga tgcaaaaacg
     1801 cgttgatggc ttgctgttac tgtgcaccga aacgcatcaa ccttcgcgtg aaatcatgca
     1861 acgttatccg acagtgccta ctgtgatgat ggactgggct ccgttcgatg gcgacagcga
     1921 tcttattcag gataactcgt tgctgggcgg agacttagca acgcaatatc tgatcgataa
     1981 aggtcatacc cgtatcgcct gtattaccgg cccgctggat aaaactccgg cgcgcctgcg
     2041 gttggaaggt tatcgggcgg cgatgaaacg tgcgggtctc aacattcctg atggctatga
     2101 agtcactggt gattttgaat ttaacggcgg gtttgacgct atgcgccaac tgctatcaca
     2161 tccgctgcgt cctcaggccg tctttaccgg aaatgacgct atggctgttg gcgtttacca
     2221 ggcgttatat caggcagagt tacaggttcc gcaggatatc gcggtgattg gctatgacga
     2281 tatcgaactg gcaagcttta tgacgccacc attaaccact atccaccaac cgaaagatga
     2341 actgggggag ctggcgattg atgtactcat ccatcggata acccagccga cccttcagca
     2401 acaacgatta caacttactc cgattctgat ggaacgcggt tcggcttagt aaccatgggc
     2461 gcaacgcaat taatgtaagt tagcgcggcc ccacgggtgc gcatgatcgt gctcctgtcg
     2521 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     2581 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     2641 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     2701 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     2761 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     2821 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     2881 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     2941 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     3001 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     3061 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     3121 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3181 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3241 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     3301 actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc
     3361 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     3421 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3481 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3541 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3601 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3661 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3721 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3781 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3841 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3901 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3961 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4021 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4081 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4141 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4201 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4261 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4321 gtaaacttgg tctgacagct tggtctgaca gttacgcccc gccctgccac tcatcgcagt
     4381 actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg gcatgatgaa
     4441 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatcgtga
     4501 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     4561 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     4621 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     4681 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     4741 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cggaattccg
     4801 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     4861 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     4921 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     4981 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     5041 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     5101 ttacgtgccc gatcaactta ccaatgctgt atttagaaaa ataaacaaat aggggttccg
     5161 cgcacatttc cccgaaaagt gccacctgaa attgtaaacg ttaatatttt gttaaaattc
     5221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5281 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5401 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5641 gcgtcccatt cgcca
//
