LOCUS       sequence-296715-        3592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T3 promoter driven synthesis of TetR-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3592)
  AUTHORS   Niederholtmeyer H, Chaggan C, Devaraj NK
  TITLE     Communication and quorum sensing in non-living mimics of eukaryotic 
            cells.
  JOURNAL   Nat Commun. 2018 Nov 28;9(1):5027. doi: 10.1038/s41467-018-07473-7.
  PUBMED    30487584
REFERENCE   2  (bases 1 to 3592)
  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: "Nat 
            Commun."; date: "2018-11-28"; volume: "9(1):5027. doi"; pages: " 
            10.1038/s41467-018-07473-7"
COMMENT     Sequence Label: pSB1C3-pT3-tetR-sfGFP
FEATURES             Location/Qualifiers
     source          1..3592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             94..714
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(167..186)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             715..1428
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1457..1504
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1550..1593
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1597..1615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1777..2436
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1843..1862)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2457..2551
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2734..3322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3223..3242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3476..3493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3517..3574)
                     /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."
ORIGIN
        1 gtcttcacct cgaggatctt aaggctagag aattaacact cactaaaggg agatgtggtc
       61 tagacattcc aggttaagaa ggaggaaaaa aaaatgtcca gattagataa aagtaaagtg
      121 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
      181 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
      241 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
      301 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
      361 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
      421 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
      481 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
      541 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
      601 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
      661 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtctatgcgt
      721 aaaggagaag aacttttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc
      781 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
      841 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
      901 acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac
      961 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
     1021 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
     1081 attgagctga aaggcattga ctttaaagaa gatggcaata tcctgggcca taagctggaa
     1141 tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa
     1201 gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac
     1261 cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc
     1321 acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag
     1381 ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaata ataacgactc
     1441 aggctgctac tcaaaactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1501 tttgggcttg atatcgaatt cctgcagccc ggggcggccg cgaattccag aaatcatcct
     1561 tagcgaaagc taaggatttt ttttatctga aattctgcct cgtgatacgc ctatttttat
     1621 aggttaatgt catgataata atggtttctt agacgtcagg tggcactcga gttgatcggg
     1681 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1741 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1801 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1861 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1921 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     1981 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2041 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     2101 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     2161 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     2221 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2281 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2341 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2401 caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact
     2461 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     2521 ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc gagctgtcag accaagttta
     2581 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2641 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3301 aggggggcgg agcctgtgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3361 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3421 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3481 gtcagtgagc gaggaagcct gcataacgcg aagtaatctt ttcggtttta aagaaaaagg
     3541 gcagggtggt gacaccttgc ccttttttgc cggactgcag cggccgctac ta
//
