LOCUS       sequence-416180-        2039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of polyProline FETCH reporter 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-p5-PRB4_R7-9-FETCH1
FEATURES             Location/Qualifiers
     source          1..2039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        28..46
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             78..100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             354..371
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(435..453)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      449..496
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(514..1329)
                     /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     1237..1256
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1321..1340
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1451..2039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1940..1959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt
       61 ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg gactacaaag
      121 acgaagacct gctgccgccc cctcctccta tcccaaagcc tgaaggacga ccccctcagg
      181 gtgggaacca atcgcaggga ccacccccgc atcccggaaa accagagaga ccaccgcctc
      241 aaggtggcaa tcagtctcac agaccacccc caccacccgg aaaacctgaa agacccccac
      301 ctcaaggcgg taaccaatcc caaggtccgc cccctcatcc gggcttttta aactgctgtc
      361 ctgggtgttg tatggaaccg taagtcgacc ggctgctaac aaagcccgaa aggaagctga
      421 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      481 cttgaggggt tttttgctga aagccaattc tgattagaaa aactcatcga gcatcaaatg
      541 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
      601 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
      661 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
      721 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
      781 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
      841 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
      901 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
      961 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1021 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1081 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1141 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1201 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1261 taaatcagca tccatgttgg aatttaatcg cggcttcgag caagacgttt cccgttgaat
     1321 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     1381 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtgagatca
     1441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaa
//
