LOCUS       sequence-197263-        3980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sigma 54 protein in E. coli. The expression is controlled 
            by a OR2-OR1-Pr promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3980)
  AUTHORS   Guo S, Yeung E, Murray RM
  TITLE     Implementation and System Identification of a Phosphorylation-Based 
            Insulator in a Cell-Free Transcription-Translation System
  JOURNAL   bioRxiv 122606
REFERENCE   2  (bases 1 to 3980)
  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: "bioRxiv 
            122606"
COMMENT     Sequence Label: pSG09-OR2-OR1-Pr-sigma54
FEATURES             Location/Qualifiers
     source          1..3980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(166..270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     338..356
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             723..743
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     rep_origin      2373..2918
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2819..2838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3035..3052
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(3285..3980,1..96,97..165))
                     /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     3908..3927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
       61 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      301 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
      361 tcgtcttcaa gaattctggc gaatcctctg accagccaga aaacgacctt tctgtggtga
      421 aaccggatgc tgcaattcag agcgccagca agtgggggac agcagaagac ctgaccgccg
      481 cagagtggat gtttgacatg gtgaagacta tcgcaccatc agccagaaaa ccgaattttg
      541 ctgggtgggc taacgatatc cgcctgatgc gtgaacgtga cggacgtaac caccgcgaca
      601 tgtgtgtgct gttccgctgg gcatgccagg acaacttctg gtccggtaac gtgctgagct
      661 aacaccgtgc gtgttgacaa ttttacctct ggcggtgata atggttgcag ctagcaataa
      721 ttttgtttaa ctttaagaag gaggatccaa atgaagcaag gtttgcaact caggcttagc
      781 caacaactgg cgatgacgcc acagctccaa caggcaattc gtctgttgca gttgtcgacg
      841 ctggaacttc agcaggagct acagcaggcg ctggagagta atccgctgct tgagcaaatc
      901 gacactcatg aagaaatcga cacccgcgaa acgcaagaca gtgaaacgct ggacaccgcc
      961 gacgcgctcg aacaaaaaga gatgccggaa gagctgccgc tcgatgccag ttgggacacc
     1021 atttacaccg ctggtacacc atccggcacc agcggtgact acattgacga cgagctgccg
     1081 gtctaccagg gcgaaacgac gcagaccttg caggattacc tgatgtggca ggttgagctg
     1141 acaccgtttt ccgacactga ccgcgctatt gctacctcta tcgtcgatgc cgttgatgaa
     1201 accggttatc tgactgtccc gctggaagat attctcgaaa gtataggcga tgaagagatc
     1261 gacatcgacg aagttgaagc cgtccttaag cggatccaac ggtttgatcc ggtcggtgtg
     1321 gcggcaaaag atctgcgtga ctgtctgctg atccaactct cccaattcga taagaccacg
     1381 ccgtggctgg aagaggccag actgatcatt agcgatcatc tcgatctgtt agccaatcac
     1441 gacttccgca ctttaatgcg cgtcacgcgt ctgaaagaag atgtgctgaa agaagccgtc
     1501 aatctgatcc agtcgctcga tccgcgcccc gggcagtcga tccagactgg cgaacctgag
     1561 tatgtcattc cagatgtgct ggtgcgtaag cataacggtc actggacggt agaactcaac
     1621 agtgacagca ttccgcgtct gcaaatcaac cagcactacg cctcgatgtg caataacgcg
     1681 cgcaacgatg gtgacagcca gtttatccgc agcaatctgc aggatgccaa atggttgatc
     1741 aagagtctgg aaagccgtaa cgatacgcta ctgcgcgtga gtcgctgtat cgttgaacag
     1801 cagcaagcct tctttgagca aggtgaagaa tatatgaaac cgatggtact ggccgatatc
     1861 gcccaggctg tcgaaatgca tgaatcgacg atatctcgcg tgaccacgca aaaatacctg
     1921 catagtccac gaggcatttt tgaactgaag tatttctttt ccagtcacgt caataccgag
     1981 ggcggcggcg aagcttcctc cacggcgatt cgtgcgctgg tgaagaaatt aatcgcggcg
     2041 gaaaacccag cgaaaccgtt gagcgacagc aagttaacct ctttgctgtc ggaacaaggt
     2101 atcatggtgg cacgccgcac tgttgcgaag taccgagagt ctttatccat tccgccgtca
     2161 aaccagcgta aacaactcgt ttgatctaga ggcgccactc gagagtcgac caaagcccgc
     2221 cgaaaggcgg gcttttctgt gccggcatga taagctgtca aacatgagaa ttgcaactta
     2281 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag
     2341 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     2401 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     2461 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     2521 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     2581 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     2641 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     2701 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     2761 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     2821 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     2881 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     2941 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     3001 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     3061 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     3121 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     3181 agggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3241 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3301 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3361 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3421 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3481 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3541 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3601 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3661 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3721 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3781 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3841 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3901 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3961 tcttcggggc gaaaactctc
//
