LOCUS       sequence-275232-        4672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PDNsp1 - TARGET8 - sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4672)
  AUTHORS   Glasscock CJ, Biggs BW, Lazar JT, Arnold JH, Burdette LA, Valdes A, 
            Kang MK, Tullman-Ercek D, Tyo KEJ, Lucks JB
  TITLE     Dynamic Control of Gene Expression with Riboregulated Switchable 
            Feedback Promoters.
  JOURNAL   ACS Synth Biol. 2021 May 21;10(5):1199-1213. doi: 
            10.1021/acssynbio.1c00015. Epub 2021 Apr 9.
  PUBMED    33834762
REFERENCE   2  (bases 1 to 4672)
  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.1c00015"; journalName: "ACS Synth Biol"; date: 
            "2021-05-21- 21"; volume: "10"; issue: "5"; pages: "1199-1213"
COMMENT     Sequence Label: pJBL6721
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..95)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(116..775)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     690..709
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     937..955
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(959..1002)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1005..1026
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(1308..1310,1311..2024)
                     /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             1308..2021
                     /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(1338..1366)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1961..1981
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2041..2112
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2128..2155
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2174..2194
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2195..2252
                     /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(2276..2293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      2827..3049
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3097..4047
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
       61 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
      121 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
      181 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
      241 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
      301 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
      361 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
      421 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
      481 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
      541 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
      601 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
      661 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
      721 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
      781 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
      841 cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc
      901 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
      961 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
     1021 ctagaggaag acacggagtt cagccaaaaa acttaagacc gccggtcttg tccactacct
     1081 tgcagtaatg cggtggacag gatcggcggt tttcttttct cttctcaagc ttggcgcgcc
     1141 tcagtcagag tattgactta taactcaacc tataggattc ttacagccat cgagagctgc
     1201 gccatcctca atctctacct actctcactt actcttatcc tgcggggaat gtatacagtt
     1261 catgtatata ttccccgctt tttttttgga tctaggagga aggatctatg agcaaaggag
     1321 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     1381 acaaattttc tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta
     1441 aatttatttg cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga
     1501 cctatggtgt tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca
     1561 agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga
     1621 cctacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt
     1681 taaagggtat tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact
     1741 ttaactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact
     1801 tcaaaattcg ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa
     1861 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat
     1921 ctgtcctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa
     1981 ctgctgctgg gattacacat ggcatggatg agctctacaa ataatactag agccaggcat
     2041 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2101 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     2161 acgctatgtc ttctactagt agcggccgct gcagtccgac aaaaaagggc aaggtgtcac
     2221 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc
     2281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2341 cggtaatgac agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt
     2401 agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg
     2461 gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac
     2521 gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc
     2581 ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga
     2641 actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata
     2701 ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg
     2761 ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa
     2821 agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt
     2881 attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa
     2941 tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag
     3001 atacccacaa ctcaaaggaa aaggactagt aattatcatt gactagccca tctcaattgg
     3061 tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa
     3121 gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta
     3181 tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta
     3241 tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt
     3301 gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg
     3361 gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta
     3421 gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa
     3481 tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg
     3541 ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat
     3601 gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa
     3661 tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt
     3721 gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg
     3781 gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca
     3841 accattacat cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta
     3901 actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag
     3961 tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac
     4021 atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc
     4081 atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact
     4141 gtccatatgc acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt
     4201 ttttggtgca ttcaaagctg ttcaccatga acagatcgac aatgtaacag atgaacagca
     4261 tgtaacacct aatagaacag gtgaaaccag taaaacaaag caactagaac atgaaattga
     4321 acacctgaga caacttgtta cagctcaaca gtcacacata gacagcctga aacaggcgat
     4381 gctgcttatc gaatcaaagc tgccgacaac acgggagcca gtgacgcctc ccgtggggaa
     4441 aaaatcatgg caattctgga agaaatagcg ctttcagccg gcaaaccggc tgaagccgga
     4501 tctgcgattc tgataacaaa ctagcaacac cagaacagcc cgtttgcggg cagcaaaacc
     4561 cgtaccgatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4621 aatcaatcta aagtatatat gagtaaactt ggtctgacag ctcgaggctt gg
//
