LOCUS       sequence-132917-        4844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET pduD1-224.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4844)
  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 4844)
  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: EYK346
FEATURES             Location/Qualifiers
     source          1..4844
                     /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      1432..1503
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1519..1546
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2077..3027)
                     /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(3789..3883)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3914..4708)
                     /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(4025..4044)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4635..4654
                     /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 atggaaatta atgaaaaatt gctgcgccag ataattgaag acgtactccg
      781 cgatatgaag 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 gaccaagtac gtcgtgacgg gcaaaaaccc
     1381 gcaggaactg cgcgtggcgc tttaataact cgagtaagga tctccaggca tcaaataaaa
     1441 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1501 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacctagggt
     1561 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
     1621 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
     1681 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
     1741 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     1801 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     1861 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     1921 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     1981 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     2041 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     2101 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     2161 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     2221 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     2281 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     2341 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     2401 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     2461 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     2521 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     2581 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     2641 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     2701 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     2761 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     2821 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     2881 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     2941 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     3001 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     3061 ataccaattg agatgggcta gtcaatgata attactagtc cttttcccgg gtgatctggg
     3121 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     3181 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     3241 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     3301 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     3361 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     3421 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     3481 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     3541 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     3601 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     3661 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
     3721 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta
     3781 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     3841 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctctcgaac
     3901 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3961 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     4021 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     4081 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     4141 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     4201 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     4261 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     4321 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     4381 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     4441 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4501 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     4561 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4621 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4681 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     4741 atcatgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     4801 agggcttccc aaccttacca gagggcgccc cagctggcaa ttcc
//
