LOCUS       sequence-416177-        2039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of polyProline FETCH control from the T7 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2039)
  AUTHORS   Willi JA, Karim AS, Jewett MC
  TITLE     Cell-Free Translation Quantification via a Fluorescent Minihelix.
  JOURNAL   ACS Synth Biol. 2024 Jul 19;13(7):2253-2259. doi: 
            10.1021/acssynbio.4c00266. Epub 2024 Jul 9.
  PUBMED    38979618
REFERENCE   2  (bases 1 to 2039)
  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/acssynbio.4c00266"; journalName: "ACS Synth Biol"; date: 
            "2024-07-19- 19"; volume: "13"; issue: "7"; pages: "2253-2259"
COMMENT     Sequence Label: pJL1-sFLAG-a5-PRB4_R7-9A-FETCH1
FEATURES             Location/Qualifiers
     source          1..2039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             247..264
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(328..346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      342..389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(407..1222)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1130..1149
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1214..1233
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1344..1932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1833..1852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1960..1979
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1960..1978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2010..2032
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atggactaca aagacgaaga cctgctggcc gctgcagctg caattgccaa ggccgagggt
       61 cgtgcggctc agggcgggaa tcaaagtcag ggtgcagctg cacatgcagg taaagccgag
      121 cgcgcagccg ctcaaggcgg taatcagtca caccgtgccg ctgccgcagc tggtaaagca
      181 gaacgagctg ccgctcaagg tggaaaccaa tctcaaggag cggcggctca tgctggtttt
      241 ttaaactgct gtcctgggtg ttgtatggaa ccgtaagtcg accggctgct aacaaagccc
      301 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      361 cctctaaacg ggtcttgagg ggttttttgc tgaaagccaa ttctgattag aaaaactcat
      421 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
      481 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
      541 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
      601 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
      661 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
      721 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
      781 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
      841 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
      901 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
      961 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     1021 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     1081 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     1141 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcttc gagcaagacg
     1201 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     1261 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     1321 caacgtgaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1441 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1501 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1561 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1621 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1681 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1741 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1801 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1861 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1921 agcctatgga aaaacgccag caacgcgatc ccgcgaaatt aatacgactc actataggga
     1981 gaccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacat
//
