LOCUS       sequence-193802-        4466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR protein in E. coli. The expression is controlled by a
            pBAD promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4466)
  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 4466)
  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: pSG22-pBAD-TetR
FEATURES             Location/Qualifiers
     source          1..4466
                     /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..958
                     /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      1054..1125
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1141..1168
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1699..2649)
                     /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(3411..3505)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3536..4330)
                     /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(3647..3666)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4257..4276
                     /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 ggtcttaatg
      961 aactcaaagc ccgccgaaag gcgggctttt ctgtccttga gagtcgtatg ttgtcttcta
     1021 aggatccaaa ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     1081 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1141 ggctcacctt cgggtgggcc tttctgcgtt tatacctagg gtacgggttt tgctgcccgc
     1201 aaacgggctg ttctggtgtt gctagtttgt tatcagaatc gcagatccgg cttcagccgg
     1261 tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc cacgggaggc
     1321 gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc ctgtttcagg
     1381 ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat ttcatgttct
     1441 agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg ttcatctgtt
     1501 acattgtcga tctgttcatg gtgaacagct ttgaatgcac caaaaactcg taaaagctct
     1561 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     1621 atgtaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     1681 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     1741 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatacttac tttgcatgtc
     1801 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     1861 tgtttttctt agtccgttat gtaggtagga atctgatgta atggttgttg gtattttgtc
     1921 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     1981 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     2041 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     2101 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     2161 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     2221 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     2281 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     2341 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     2401 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     2461 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     2521 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     2581 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     2641 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     2701 tagtcaatga taattactag tccttttccc gggtgatctg ggtatctgta aattctgcta
     2761 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     2821 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     2881 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     2941 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     3001 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     3061 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     3121 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     3181 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     3241 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     3301 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     3361 gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca
     3421 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3481 attactggat ctatcaacag gagtccaagc gagctctcga accccagagt cccgctcaga
     3541 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     3601 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     3661 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     3721 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     3781 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     3841 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     3901 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     3961 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4021 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4081 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4141 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4201 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4261 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4321 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcatgat cccctgcgcc
     4381 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     4441 cagagggcgc cccagctggc aattcc
//
