LOCUS       sequence-132913-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET etuAld+Ile1-20-pduD21-224- FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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: CMJ162
FEATURES             Location/Qualifiers
     source          1..4873
                     /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)"
     CDS             1406..1429
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1461..1532
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1548..1575
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2106..3056)
                     /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(3818..3912)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3943..4737)
                     /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(4054..4073)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4664..4683
                     /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 atggataagg acttacagtc aatacaggaa attgtaagaa ctcttatagc
      781 aaaagcaaag aaaggcagcg ataaacccgt ctcgtttaat gcgcctgcgg catccacagc
      841 accacagacc gctgcgcctg cgggcgacgg ctttctgacc gaagtgggcg aagcgcgcca
      901 gggcactcag caggacgaag tcattatcgc cgtcggcccg gcatttggcc tggcgcaaac
      961 cgtcaatatc gtcggcttac cgcataagag cattctgcgc gaagtcattg ccggtattga
     1021 agaagaaggc atcaaggcgc gcgtgattcg ctgctttaaa tcttccgacg tggcgttcgt
     1081 cgccgttgaa ggtaaccgcc tgagcggatc cggcatctcc atcggcatcc agtcgaaagg
     1141 tactacggtt atccaccagc aggggctacc gccgctctcc aacctggagc tgttcccgca
     1201 ggcaccgctg ctgacgctgg aaacctaccg tcagattggt aaaaacgccg cccgctatgc
     1261 gaaacgagaa tcaccgcagc cggtccctac gctcaatgac cagatggcac gcccgaagta
     1321 ccaggcaaag tcggccattt tacatattaa agagaccaag tacgtcgtga cgggcaaaaa
     1381 cccgcaggaa ctgcgcgtgg cgcttgatta taaagatgac gatgacaagt aaaagctttc
     1441 gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1501 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     1561 gtgggccttt ctgcgtttat acctagggta cgggttttgc tgcccgcaaa cgggctgttc
     1621 tggtgttgct agtttgttat cagaatcgca gatccggctt cagccggttt gccggctgaa
     1681 agcgctattt cttccagaat tgccatgatt ttttccccac gggaggcgtc actggctccc
     1741 gtgttgtcgg cagctttgat tcgataagca gcatcgcctg tttcaggctg tctatgtgtg
     1801 actgttgagc tgtaacaagt tgtctcaggt gttcaatttc atgttctagt tgctttgttt
     1861 tactggtttc acctgttcta ttaggtgtta catgctgttc atctgttaca ttgtcgatct
     1921 gttcatggtg aacagctttg aatgcaccaa aaactcgtaa aagctctgat gtatctatct
     1981 tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatg taacggtgaa
     2041 cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca agagccataa
     2101 gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc
     2161 gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt
     2221 gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt
     2281 ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt
     2341 atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa
     2401 atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg
     2461 tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag
     2521 ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg
     2581 tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg
     2641 ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga
     2701 taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag
     2761 tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc
     2821 gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct actgatgttc
     2881 atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt
     2941 tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt
     3001 aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat
     3061 ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa
     3121 ttactagtcc ttttcccggg tgatctgggt atctgtaaat tctgctagac ctttgctgga
     3181 aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt
     3241 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga
     3301 atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg
     3361 atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt
     3421 agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca
     3481 cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa
     3541 tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat
     3601 acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc
     3661 tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg
     3721 gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat
     3781 caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc
     3841 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     3901 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
     3961 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4021 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4081 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4141 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4201 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     4261 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     4321 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     4381 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4441 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4501 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     4561 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4621 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4681 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     4741 aaacgatcct catcctgtct cttgatcaga tcatgatccc ctgcgccatc agatccttgg
     4801 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     4861 agctggcaat tcc
//
