LOCUS       sequence-256397-        4496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR protein with ssrA degradation tag. The expression is 
            controlled by a pLas promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4496)
  AUTHORS   Guo S, Murray RM
  TITLE     Construction of Incoherent Feedforward Loop Circuits in a Cell-Free 
            System and in Cells.
  JOURNAL   ACS Synth Biol. 2019 Mar 15;8(3):606-610. doi: 
            10.1021/acssynbio.8b00493. Epub 2019 Mar 6.
  PUBMED    30790525
REFERENCE   2  (bases 1 to 4496)
  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.8b00493"; journalName: "ACS Synth Biol"; date: 
            "2019-03-15- 15"; volume: "8"; issue: "3"; pages: "606-610"
COMMENT     Sequence Label: pSG84-pLas-TetRssrA
FEATURES             Location/Qualifiers
     source          1..4496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             309..929
                     /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(382..401)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1084..1155
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1171..1198
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1729..2679)
                     /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(3441..3535)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3566..4360)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3677..3696)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4287..4306
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaatgtg cattgttctc gtgtgaagcc attgctctga
      121 tcttttggac gtttcttcga gcctagcaag ggtccgggtt caccgaaatc tatctcattt
      181 gctagttata aaattatgaa atttgtataa attcttcaga agcgggccca agttcactta
      241 aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca tgcggtggag
      301 ggtttctaat gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta
      361 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
      421 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
      481 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
      541 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
      601 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
      661 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg
      721 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
      781 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
      841 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
      901 aaaaacaact taaatgtgaa agtgggtctg cagcaaacga cgaaaactac gctttagcag
      961 cttaatgaac taacgcatga gaaagccccc ggaagatcac cttccggggg cttttttatt
     1021 gcgcccttga gagtcgtatg ttgtcttcta aggatccaaa ctcgagtaag gatctccagg
     1081 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1141 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     1201 tatacctagg gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt
     1261 tatcagaatc gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag
     1321 aattgccatg attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt
     1381 gattcgataa gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca
     1441 agttgtctca ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt
     1501 ctattaggtg ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct
     1561 ttgaatgcac caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca
     1621 tctgtgcata tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg
     1681 tttgttagtc ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc
     1741 gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga
     1801 accattgaga tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag
     1861 ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga
     1921 atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag
     1981 ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg
     2041 gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat
     2101 tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa
     2161 catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt
     2221 tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac
     2281 atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc
     2341 actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat
     2401 ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt
     2461 tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg
     2521 gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag
     2581 tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat
     2641 cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg
     2701 attttaatca ctataccaat tgagatgggc tagtcaatga taattactag tccttttccc
     2761 gggtgatctg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
     2821 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
     2881 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
     2941 gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt
     3001 tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc
     3061 gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt
     3121 ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact
     3181 acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca
     3241 cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg
     3301 ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg
     3361 tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc
     3421 ttactgtccc tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     3481 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     3541 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     3601 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     3661 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     3721 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     3781 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     3841 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     3901 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     3961 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4021 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4081 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4141 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     4201 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4261 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4321 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     4381 tctcttgatc agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     4441 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattcc
//
