LOCUS       sequence-182441-        3440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR in the presence of ipgC and mxiE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  TITLE     Unpublished Murray Lab Plasmids
REFERENCE   2  (bases 1 to 3440)
  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"
COMMENT     Sequence Label: PipaH-TetR
FEATURES             Location/Qualifiers
     source          1..3440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             222..244
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             251..874
                     /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(324..343)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1062..1079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1233..1821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1313..1332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1992..2783,2784..2852))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2615..2634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2853..2957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3025..3043
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcgatctgt tgccttggaa agttgatctg acctctcagt aaatatcaat acggttctga
       61 cgagccgctt accgttcaaa tatgaagtac gatgtttaac taaccgaaaa acaagaacaa
      121 tacggtgcaa acaggccatt cacggttaac tgaaacagta tcgttttttt acagccaatt
      181 ttgtttatcc ttattaagat aaaaaagtgc taagcaataa ttttgtttaa ctttaagaag
      241 gagatataca atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct
      301 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga
      361 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat
      421 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga
      481 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc
      541 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt
      601 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt
      661 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga
      721 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt
      781 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt
      841 agaaaaacaa cttaaatgtg aaagtgggtc ttaatgaact aacgcatgag aaagcccccg
      901 gaagatcacc ttccgggggc ttttttattg cgcccttgag agtcgggcat tgtcttcgct
      961 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtg ttctttatca
     1021 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     1081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1141 tcaacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1381 ggatacctgt ccgcctttct ccattcggga agcgtggcgc tttctcatag ctcacgctgt
     1441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2101 ccatctggcc ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta
     2161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttgcatgatc ccccatgttg
     2401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2581 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     2641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3001 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tgttcaagaa
     3061 ttctggcgaa tcctctgacc agccagaaaa cgacctttct gtggtgaaac cggatgctgc
     3121 aattcagagc gccagcaagt gggggacagc agatgacctg accgccgcag agtggatgtt
     3181 tgacatggtg atgactatcg caccatcagc cagaaaaccg aattttgctg ggtgggctaa
     3241 cgatatccgc ctgatgcgtg aacgtgacgg acgtaacgaa gacaaccacg cattgctgtt
     3301 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     3361 tagggtcaaa aatcgtggcg ttgacaaaat ggctgcgtta cgtcattgag catatccagg
     3421 actggccggc aaaccgggta
//
