LOCUS       sequence-108023-        5363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OmpR constitutively from PompB97 and sfGFP under the 
            PompF146 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5363)
  AUTHORS   Schmidl SR, Sheth RU, Wu A, Tabor JJ
  TITLE     Refactoring and optimization of light-switchable escherichia coli 
            two-component systems.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):820-31. doi: 10.1021/sb500273n. 
            Epub 2014 Oct 11.
  PUBMED    25250630
REFERENCE   2  (bases 1 to 5363)
  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/sb500273n";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "820-31"
COMMENT     Sequence Label: pSR59.4
FEATURES             Location/Qualifiers
     source          1..5363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             147..860
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      881..952
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      968..995
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1949..2020
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2036..2063
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2070..2127
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2151..2168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2430..3380)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3428..3650
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4142..4236)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4266..5057,5058..5126))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHLRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4889..4908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5127..5231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5296..5314
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5318..5361)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gttccttaaa ttttactttt ggttacatat tttttctttt tgaaaccaaa tctttatctt
       61 tgtagcactt tcacggtagc gaaacgttag tttgaatgga aagatgcctg caggtgtcat
      121 aaaaaaaacc atgagggtaa taaataatgc gtaaaggcga agagctgttc actggtgtcg
      181 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
      241 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
      301 aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
      361 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
      421 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
      481 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
      541 aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
      601 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
      661 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
      721 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
      781 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
      841 gtatggatga actgtacaaa tgatgaagat ctccaggcat caaataaaac gaaaggctca
      901 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
      961 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat agtaaatcac tgcataattc
     1021 gtgtcgatta aaccgcctaa gtcccccagg aaagggggat ataacagtat agattttgtc
     1081 agccttcagc ttggctttac cgtcaaaaaa ataggctgaa attcatacca gatttagctg
     1141 gtgacgaacg tgagcttttt taagaataca cgcttacaaa ttgttgcgaa cctttgggag
     1201 tacaaacaat gcaagagaac tacaagattc tggtggtcga tgacgacatg cgcctgcgtg
     1261 cgctgctgga acgttatctc accgaacaag gcttccaggt tcgaagcgtc gctaatgcag
     1321 aacagatgga tcgcctgctg actcgtgaat ctttccatct tatggtactg gatttaatgt
     1381 tacctggtga agatggcttg tcgatttgcc gacgtcttcg tagtcagagc aacccgatgc
     1441 cgatcattat ggtgacggcg aaaggggaag aagtggaccg tatcgtaggc ctggagattg
     1501 gcgctgacga ctacattcca aaaccgttta acccgcgtga actgctggcc cgtatccgtg
     1561 cggtgctgcg tcgtcaggcg aacgaactgc caggcgcacc gtcacaggaa gaggcggtaa
     1621 ttgctttcgg taagttcaaa cttaacctcg gtacgcgcga aatgttccgc gaagacgagc
     1681 cgatgccgct caccagcggt gagtttgcgg tactgaaggc actggtcagc catccgcgtg
     1741 agccgctctc ccgcgataag ctgatgaacc ttgcccgtgg tcgtgaatat tccgcaatgg
     1801 aacgctccat cgacgtgcag atttcgcgtc tgcgccgcat ggtggaagaa gatccagcgc
     1861 atccgcgtta cattcagacc gtctggggtc tgggctacgt ctttgtaccg gacggctcta
     1921 aagcatgata ataatctaga ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1981 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     2041 accttcgggt gggcctttct gcgtttatat ccggcaaaaa agggcaaggt gtcaccaccc
     2101 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atctcgaggt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     2281 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     2341 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     2401 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     2461 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatgctta
     2521 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     2581 gcatcgtgta gtgtttttct tagtccgtta cgtaggtagg aatctgatgt aatggttgtt
     2641 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     2701 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     2761 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     2821 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     2881 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     2941 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     3001 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     3061 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     3121 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     3181 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     3241 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     3301 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     3361 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     3421 ttgagatggg ctagtcaatg ataattacat gtccttttcc tttgagttgt gggtatctgt
     3481 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     3541 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     3601 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     3661 tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga
     3721 ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt
     3781 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     3841 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     3901 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     3961 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     4021 tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg
     4081 cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg
     4141 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     4201 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct
     4261 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4321 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4381 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4441 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4501 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4561 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4621 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4681 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4741 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4801 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4861 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4921 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4981 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5041 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5101 gcgacacgta aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5161 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5221 ggggttccgc gcacatttcc catggtgcca cctgacgtct aagaaaccat tattatcatg
     5281 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     5341 gctttcgcta aggatgattt ctg
//
