LOCUS       sequence-419668-        2027 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 2027)
  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 2027)
  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-p1-PRB4_R7-9-FETCH1
FEATURES             Location/Qualifiers
     source          1..2027
                     /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             342..359
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(423..441)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      437..484
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(502..1317)
                     /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     1225..1244
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1309..1328
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1439..2027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1928..1947
                     /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 gctgccgatc ccaaagcctg aaggacgacc ccctcagggt gggaaccaat
      181 cgcagggacc acccccgcat cccggaaaac cagagagacc accgcctcaa ggtggcaatc
      241 agtctcacag accaccccca ccacccggaa aacctgaaag acccccacct caaggcggta
      301 accaatccca aggtccgccc cctcatccgg gctttttaaa ctgctgtcct gggtgttgta
      361 tggaaccgta agtcgaccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
      421 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
      481 tttgctgaaa gccaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
      541 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
      601 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
      661 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
      721 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
      781 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
      841 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
      901 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
      961 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     1021 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1081 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1141 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     1201 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1261 catgttggaa tttaatcgcg gcttcgagca agacgtttcc cgttgaatat ggctcataac
     1321 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     1381 atcttgtgca atgtaacatc agagattttg agacacaacg tgagatcaaa ggatcttctt
     1441 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1501 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1561 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1621 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1681 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1741 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1801 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1861 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1921 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1981 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaa
//
