LOCUS       sequence-122802-        4846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET eutC1-20-pduD21-224-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  AUTHORS   Jakobson CM, Kim EY, Slininger MF, Chien A, Tullman-Ercek D
  TITLE     Localization of Proteins to the 1,2-Propanediol Utilization 
            Microcompartment by Non-native Signal Sequences Is Mediated by a 
            Common Hydrophobic Motif.
  JOURNAL   J Biol Chem. 2015 Aug 17. pii: jbc.M115.651919.
  PUBMED    26283792
REFERENCE   2  (bases 1 to 4846)
  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: "J Biol 
            Chem. 2015 Aug 17. pii: jbc.M115.651919."
COMMENT     Sequence Label: CMJ156
FEATURES             Location/Qualifiers
     source          1..4846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /codon_start=1
                     /gene="tetR"
                     /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="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             715..726
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1434..1505
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1521..1548
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2079..3029)
                     /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(3791..3885)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3916..4710)
                     /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(4027..4046)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4637..4656
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcttaaagag
      721 gagaaaggtc atggatcaaa aacagattga agaaattgta cgtagcgtga tggcgtcaat
      781 gggacaggac ggcagcgata aacccgtctc gtttaatgcg cctgcggcat ccacagcacc
      841 acagaccgct gcgcctgcgg gcgacggctt tctgaccgaa gtgggcgaag cgcgccaggg
      901 cactcagcag gacgaagtca ttatcgccgt cggcccggca tttggcctgg cgcaaaccgt
      961 caatatcgtc ggcttaccgc ataagagcat tctgcgcgaa gtcattgccg gtattgaaga
     1021 agaaggcatc aaggcgcgcg tgattcgctg ctttaaatct tccgacgtgg cgttcgtcgc
     1081 cgttgaaggt aaccgcctga gcggatccgg catctccatc ggcatccagt cgaaaggtac
     1141 tacggttatc caccagcagg ggctaccgcc gctctccaac ctggagctgt tcccgcaggc
     1201 accgctgctg acgctggaaa cctaccgtca gattggtaaa aacgccgccc gctatgcgaa
     1261 acgagaatca ccgcagccgg tccctacgct caatgaccag atggcacgcc cgaagtacca
     1321 ggcaaagtcg gccattttgc atattaaaga aaccaagtac gtcgtgacgg gcaaaaaccc
     1381 gcaggaactg cgcgtggcgc tttgaaagct ttcgagtaag gatctccagg catcaaataa
     1441 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1501 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacctagg
     1561 gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc
     1621 gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag aattgccatg
     1681 attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa
     1741 gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca
     1801 ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg
     1861 ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttgaatgcac
     1921 caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata
     1981 tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg tttgttagtc
     2041 ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc
     2101 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     2161 tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     2221 gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga atctgatgta
     2281 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     2341 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     2401 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     2461 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     2521 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     2581 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     2641 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     2701 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     2761 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     2821 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     2881 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     2941 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     3001 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     3061 ctataccaat tgagatgggc tagtcaatga taattactag tccttttccc gggtgatctg
     3121 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     3181 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     3241 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     3301 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     3361 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     3421 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     3481 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     3541 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     3601 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     3661 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     3721 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc
     3781 tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     3841 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctctcga
     3901 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     3961 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     4021 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     4081 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     4141 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     4201 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     4261 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     4321 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     4381 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     4441 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     4501 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     4561 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     4621 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     4681 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     4741 agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     4801 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattcc
//
