LOCUS       sequence-275270-        7556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pstabilized - TARGET8 - sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7556)
  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 7556)
  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: pJBL6722
FEATURES             Location/Qualifiers
     source          1..7556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(3188..3190,3191..3904)
                     /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             3188..3901
                     /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(3218..3246)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3841..3861
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3921..3992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4008..4035
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4054..4074
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4075..4132
                     /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(4156..4173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4707..4929
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4977..5927
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(6553..6647)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6668..7327)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7242..7261
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7489..7507
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7511..7554)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag aggaagacac ggagctcggt accaaattcc agaaaagagg
       61 cctcccgaaa ggggggcctt ttttcgtttt ggtcctactg gcgcgcctca gtcagagtat
      121 tgacttaaag tctaacctat aggaccgtta cagccatcga gagctgcgag actgtcgccg
      181 gatgtgtatc cgacctgacg atggcccaaa agggccgaaa cagtcctcta caaataattt
      241 tgtttaaatc aattcacctg cgtgaaaatg gtagatttaa gaactttagg atattcacag
      301 cagcaacagg aaaagatcaa gcccaaagtt aggtcgacag tcgcgcagca tcacgaagcg
      361 ctggttggtc atgggtttac acatgcccac atcgtagcct tatcgcagca ccctgcagcc
      421 cttggcacgg tcgccgtcaa gtaccaggac atgattgcgg cgttgccgga agccacacat
      481 gaggcgatcg tcggtgtggg gaaacagtgg agcggagccc gagcgcttga ggccctgttg
      541 acggtcgcgg gagagctgag agggcctccc cttcagctgg acacgggcca gttgctgaag
      601 atcgcgaagc ggggaggagt cacggcggtc gaggcggtgc acgcgtggcg caatgcgctc
      661 acgggagcac ccctcaatct gaccccggat caggtggttg cgatcgccag taatggcgga
      721 ggcaaacaag cactcgaaac tgtacagcgc ctcctgccgg tactgtgcca agatcatggc
      781 ttgacgcctg agcaagtcgt agctattgca tcaaacatcg gtggtaaaca ggcgctggaa
      841 accgtacaac gattactccc tgtcttatgc caggcacacg gtctgacccc cgaccaagtt
      901 gtagccattg cgtctaatgg cggcgggaaa caggccctgg aaacggtcca acgtctgtta
      961 cccgttctgt gtcaggctca cggtctgacc cctgcccagg tagttgcaat tgccagcaac
     1021 atcggcggga aacaagcgct ggagactgtg cagcgtctgc tccctgtgtt atgccaagat
     1081 catgggctca ctccggatca ggtggtggcc atcgcttcca atattggcgg taaacaggcg
     1141 ctggagacag tgcaacgact tttacctgtt ctctgccagg atcatggtct aactcccgag
     1201 caggtcgtcg ccatcgcctc tcatgacggc gggaaacaag cgttggaaac tgtccagcga
     1261 ctcctgccgg ttttgtgcca ggcccacggg cttactcctg accaggtagt tgcgatcgcg
     1321 tcaaatgggg gtggcaaaca agccctcgaa accgtgcaac gcctgctgcc cgtcttgtgc
     1381 caagctcatg ggctgactcc ggcgcaagta gtcgcgattg cgagccacga tggcggtaag
     1441 caggcactgg aaacggttca gcgcctgctc ccggttctat gccaggatca cggcctgacc
     1501 ccggaccagg tcgtcgcgat cgcgtcaaat atcggtggca aacaagcttt ggagacagta
     1561 cagcgcctgt taccagtgct ttgccaggac catggtctga cccctgagca agtagtggcg
     1621 atcgcttcta atattggggg caaacaagcg ctggaaacag tacagcgtct gttaccggtc
     1681 ctatgccagg cacatggcct gacccctgat caggtggtag ccattgccag tcatgatggc
     1741 ggtaaacagg cgcttgagac tgtccaacgt ctgctgccgg tcctctgtca ggctcatggc
     1801 ctgacgccag ctcaagtcgt ggctatcgct tcgcatgatg gcggaaaaca ggcactggag
     1861 actgtgcagc gactgttgcc agttctgtgt caggatcacg gtttaactcc ggaccaggtg
     1921 gtcgctattg cgtcgaatgg gggcggtaaa caagcgctgg aaactgtgca acgtttgctc
     1981 ccagttctgt gccaggacca tgggctgact ccggaacagg tagtggccat tgcttctaat
     2041 attggtggga aacaggcgct ggaaaccgtg cagcgcctgc ttccagtgct ttgccaggcc
     2101 catggcctga cgccagatca ggtggttgct atagccagca atggcggcgg taaacaggcc
     2161 ctcgaaaccg tccagcgcct gctccctgtg ctgtgccagg cccatgggct taccccagcg
     2221 caagtagtgg cgattgcgtc taatattggt ggtaaacagg cgttggagac tgtacaacgc
     2281 ctgctgccag ttttatgcca agatcatggt ctgacccctg agcaggtagt ggctattgca
     2341 tccaacaacg ggggcagacc cgcactggag tcaatcgtgg cccagctttc gaggccggac
     2401 cccgcgctgg ccgcactcac taatgatcat cttgtagcgc tggcctgcct cggcggacga
     2461 cccgccttgg atgcggtgaa gaaggggctc ccgcacgcgc ctgcattgat taagcggacc
     2521 aacagaagga ttcccgagag gacatcacat cgagtggcag atcacgcgca agtggtccgc
     2581 gtgctcggat tcttccagtg tcactcccac cccgcacaag cgttcgatga cgccatgact
     2641 caatttggta tgtcgagaca cggactgctg cagctctttc gtagagtcgg tgtcacagaa
     2701 ctcgaggccc gctcgggcac actgcctccc gcctcccagc ggtgggacag gattctccaa
     2761 gcgagcggta tgaaacgcgc gaagccttca cctacgtcaa ctcagacacc tgaccaggcg
     2821 agccttcatg cgttcgcaga ctcgctggag agggatttgg acgcgccctc gcccatgcat
     2881 gaaggggacc aaactcgcgc gtcataatag gtttcagcca aaaaacttaa gaccgccggt
     2941 cttgtccact accttgcagt aatgcggtgg acaggatcgg cggttttctt ttctcttctc
     3001 aagcttatcc cgaaaattta tcaaaaagag tattgactta taactcaacc tataggatac
     3061 ttacagccat cgagagctgc gccatcctca atctctacct actctcactt actcttatcc
     3121 tgcggggaat gtatacagtt catgtatata ttccccgctt tttttttgga tctaggagga
     3181 aggatctatg agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     3241 agatggtgat gttaatgggc acaaattttc tgtccgtgga gagggtgaag gtgatgctac
     3301 aaacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg ttccgtggcc
     3361 aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc cggatcacat
     3421 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
     3481 atctttcaaa gatgacggga cctacaagac gcgtgctgaa gtcaagtttg aaggtgatac
     3541 ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa acattcttgg
     3601 acacaaactc gagtacaact ttaactcaca caatgtatac atcacggcag acaaacaaaa
     3661 gaatggaatc aaagctaact tcaaaattcg ccacaacgtt gaagatggtt ccgttcaact
     3721 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     3781 ccattacctg tcgacacaat ctgtcctttc gaaagatccc aacgaaaagc gtgaccacat
     3841 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
     3901 ataatactag agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3961 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     4021 gtgggccttt ctgcgtttat acgctatgtc ttctactagt agcggccgct gcagtccggc
     4081 aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
     4141 cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4201 ggtatcagct cactcaaagg cggtaatgac agtaagacgg gtaagcctgt tgatgatacc
     4261 gctgccttac tgggtgcatt agccagtctg aatgacctgt cacgggataa tccgaagtgg
     4321 tcagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga tagcaccaca
     4381 tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg
     4441 tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc
     4501 cagagccgtg agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat
     4561 ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc
     4621 ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt
     4681 aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta
     4741 tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa
     4801 cacacaaagg tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa
     4861 aggtctagca gaatttacag atacccacaa ctcaaaggaa aaggactagt aattatcatt
     4921 gactagccca tctcaattgg tatagtgatt aaaatcacct agaccaattg agatgtatgt
     4981 ctgaattagt tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc
     5041 atgaaaccaa gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta
     5101 caaggaaaga acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca
     5161 gtagggaaaa tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg
     5221 tggaaatcag gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca
     5281 agttctcaag cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc
     5341 agttaaaaaa attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact
     5401 ctatgaggat ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa
     5461 atatagagat tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg
     5521 agtttaaaag gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca
     5581 gcaatatgaa attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag
     5641 ttgaactaga tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga
     5701 atggtgacaa aataccaaca accattacat cagattccta cctacataac ggactaagaa
     5761 aaacactaca cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt
     5821 tgagtgacat gcaaagtaag tatgatctca atggttcgtt ctcatggctc acgcaaaaac
     5881 aacgaaccac actagagaac atactggcta aatacggaag gatctgaggt tcttatggct
     5941 cttgtatcta tcagtgaagc atcaagacta acaaacaaaa gtagaacaac tgttcaccgt
     6001 tacatatcaa agggaaaact gtccatatgc acagatgaaa acggtgtaaa aaagatagat
     6061 acatcagagc ttttacgagt ttttggtgca ttcaaagctg ttcaccatga acagatcgac
     6121 aatgtaacag atgaacagca tgtaacacct aatagaacag gtgaaaccag taaaacaaag
     6181 caactagaac atgaaattga acacctgaga caacttgtta cagctcaaca gtcacacata
     6241 gacagcctga aacaggcgat gctgcttatc gaatcaaagc tgccgacaac acgggagcca
     6301 gtgacgcctc ccgtggggaa aaaatcatgg caattctgga agaaatagcg ctttcagccg
     6361 gcaaaccggc tgaagccgga tctgcgattc tgataacaaa ctagcaacac cagaacagcc
     6421 cgtttgcggg cagcaaaacc cgtaccgatt atcaaaaagg atcttcacct agatcctttt
     6481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6541 ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     6601 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     6661 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     6721 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     6781 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     6841 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     6901 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     6961 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     7021 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     7081 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     7141 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     7201 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     7261 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     7321 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     7381 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aactcgagtg
     7441 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7501 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctg
//
