LOCUS       sequence-373856-        4788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  titratable expression of sfGFP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4788)
  AUTHORS   Shilling PJ, Khananisho D, Cumming AJ, Soderstrom B, Daley DO
  TITLE     Signal amplification of araC pBAD using a standardized translation 
            initiation region.
  JOURNAL   Synth Biol (Oxf). 2022 Jul 5;7(1):ysac009. doi: 
            10.1093/synbio/ysac009. eCollection 2022.
  PUBMED    35903559
REFERENCE   2  (bases 1 to 4788)
  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"; journalName: "Synth Biol 
            (Oxf)."; date: "2022-07-5"; volume: "7(1):ysac009. doi"; pages: " 
            10.1093/synbio/ysac009. eCollection 2022"
COMMENT     Sequence Label: pBAD/HisB (TIR23)-sfGFP
FEATURES             Location/Qualifiers
     source          1..4788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             430..1143
                     /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="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             430..1140
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(457..485)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1080..1100
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1189..1206)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1359..1445
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1537..1564
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1583..1674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1675..1743,1744..2535)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1893..1912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2706..3294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3195..3214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3448..3465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3480..3620
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3546..3565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3706..3728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3884..4762)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aatattgaat gccgcgatct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagatcta gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      481 gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg tgatgctaca
      541 aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt tccgtggcca
      601 acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc ggatcacatg
      661 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata
      721 tctttcaaag atgacgggac ctacaagacg cgtgctgaag tcaagtttga aggtgatacc
      781 cttgttaatc gtatcgagtt aaaaggtatt gattttaaag aagatggaaa cattctcgga
      841 cacaaactgg agtacaactt taactcacac aatgtataca tcacggcaga caaacaaaag
      901 aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc cgttcaacta
      961 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1021 cattacctgt cgacacaatc tgttctttcg aaagatccca acgaaaagcg tgaccacatg
     1081 gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga actctacaaa
     1141 taagaattcg aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1201 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1261 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1321 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1381 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1441 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1501 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1561 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     1621 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1681 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1741 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1801 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1861 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     1921 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1981 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2041 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2101 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2161 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2221 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2821 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3481 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     3541 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3601 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3661 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3721 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
     3781 aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     3841 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
     3901 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
     3961 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
     4021 tggcgatagg catccgggta gtgctcaaaa gcagcttcgc ctgactaatg cgttggtcct
     4081 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
     4141 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
     4201 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt
     4261 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
     4321 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
     4381 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
     4441 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
     4501 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
     4561 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
     4621 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
     4681 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
     4741 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagag
//
