LOCUS       sequence-422348-        3713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Calprotectin sensing promoter with sfGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3713)
  AUTHORS   Xia JY, Hepler C, Tran P, Waldeck NJ, Bass J, Prindle A
  TITLE     Engineered calprotectin-sensing probiotics for IBD surveillance in 
            humans.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Aug 8;120(32):e2221121120. doi: 
            10.1073/pnas.2221121120. Epub 2023 Jul 31.
  PUBMED    37523538
REFERENCE   2  (bases 1 to 3713)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-08-8"; volume: "120(32):e2221121120. 
            doi"; pages: " 10.1073/pnas.2221121120. Epub 2023 Jul 31"
COMMENT     Sequence Label: ykgMO-IGS-sfGFP
FEATURES             Location/Qualifiers
     source          1..3713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..80)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      675..769
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1176..1193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1310..1855)
                     /direction=LEFT
                     /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     complement(1390..1409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2067..2101
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             2467..2478
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2485..2487,2488..3201)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2485..3198
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2515..2543)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3138..3158
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3266..3283
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3339..3356)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3509..3555
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             join(3708..3713,1..654)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
       61 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      121 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
      181 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
      241 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
      301 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
      361 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
      421 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
      481 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
      541 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
      601 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttgat
      661 atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
      721 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcctccg
      781 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
      841 aggatttatt tattatgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
      901 gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag
      961 tggcttctat ttctatctgc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg
     1021 gcaaaagcac cgccgtacat cagcgctagc ggagtgtata ctggcttact atgttggcac
     1081 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     1141 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     1201 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     1261 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     1321 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     1381 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     1441 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     1501 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     1561 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     1621 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     1681 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     1741 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     1801 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     1861 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     1921 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgtttga cagcttatca
     1981 tcgataagct tccgatggcg cgccgagagg ctttacactt tatccttagc tttcgctaag
     2041 gatgatttct ggaattctaa agatctttta cggctagctc agccctaggt attatgctag
     2101 ctactataac ggcaataaac tgttcacttc agtgatattt aaaatatgca tcctctccct
     2161 tttttgtaag taattattat atccgtggga gaggaataca cattgtcaat caatcatgct
     2221 gcaataaatc atcggccagt aaagtggaga tagcctccat tctcgaaaaa tccatactct
     2281 cagcgaaacc atcatcaatc actcatccag gcgtttatgg gagcgtcgcc aatggctgct
     2341 aacaatgcca gacttccccg ttgcggaaat tccacatccc acaaatagtc acagtgattg
     2401 ggtgttgaaa tgatccggat gagcatgtat ctttacggtt atgttataac ataacaggta
     2461 aaaatgaaag aggagaaagg atctatgagc aaaggagaag aacttttcac tggagttgtc
     2521 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt ccgtggagag
     2581 ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac tactggaaaa
     2641 ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca atgcttttcc
     2701 cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     2761 gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg tgctgaagtc
     2821 aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa
     2881 gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa tgtatacatc
     2941 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacgttgaa
     3001 gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     3061 gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa agatcccaac
     3121 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
     3181 atggatgagc tctacaaata aggatctgaa gcttgggccc taacaaaaac tcatctcaga
     3241 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc
     3301 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     3361 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     3421 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     3481 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     3541 ggcctttcgt tttatctgtt gtttgtcggt gaactggatc cttactcgag tctagactgc
     3601 agttgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     3661 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gag
//
