LOCUS       sequence-193797-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR protein with ssrA degradation tag. The expression is 
            controlled by a pBAD promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  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 4499)
  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: pSG13-pBAD-TetRssrA
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..166
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             335..955
                     /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(408..427)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1087..1158
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1174..1201
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1732..2682)
                     /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(3444..3538)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3569..4363)
                     /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(3680..3699)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4290..4309
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaatgtg cattgctgtt ccgctgggca tgcacattga
      121 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
      181 tcctacctga cgctttttat cgcaactctc tactgtttct ccataccgtt tttttgggct
      241 agcgcaagcg ggcccaagtt cacttaaaaa ggagatcaac aatgaaagca attttcgtac
      301 tgaaacatct taatcatgcc taggaagttt tctaatgtct agattagata aaagtaaagt
      361 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa
      421 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg
      481 ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc acttttgccc
      541 tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc
      601 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca
      661 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa
      721 tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca
      781 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt
      841 attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg
      901 ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtctgcagc
      961 aaacgacgaa aactacgctt tagcagctta atgaactcaa agcccgccga aaggcgggct
     1021 tttctgtcct tgagagtcgt atgttgtctt ctaaggatcc aaactcgagt aaggatctcc
     1081 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1141 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1201 gtttatacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt
     1261 tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc
     1321 cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc
     1381 tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta
     1441 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     1501 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     1561 gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     1621 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     1681 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     1741 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     1801 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     1861 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     1921 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     1981 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     2041 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     2101 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     2161 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     2221 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     2281 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     2341 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     2401 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     2461 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     2521 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     2581 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     2641 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     2701 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     2761 cccgggtgat ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     2821 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     2881 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     2941 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     3001 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     3061 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     3121 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     3181 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     3241 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     3301 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     3361 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc
     3421 gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     3481 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     3541 agcgagctct cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3601 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3661 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3721 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3781 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct
     3841 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     3901 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     3961 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4021 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4081 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4141 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac
     4201 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4261 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4321 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     4381 ctgtctcttg atcagatcat gatcccctgc gccatcagat ccttggcggc aagaaagcca
     4441 tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattcc
//
