LOCUS       sequence-125344-        4848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET eutE1-20-pduD21-224-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4848)
  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 4848)
  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: CMJ157
FEATURES             Location/Qualifiers
     source          1..4848
                     /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             717..728
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1436..1507
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1523..1550
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2081..3031)
                     /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(3793..3887)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3918..4712)
                     /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(4029..4048)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4639..4658
                     /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 agaaaagaga attcttaaag
      721 aggagaaagg tcatgaatca acaggatatt gaacaggtgg tgaaagcggt actgctgaaa
      781 atgaaagaca gcggcagcga taaacccgtc tcgtttaatg cgcctgcggc atccacagca
      841 ccacagaccg ctgcgcctgc gggcgacggc tttctgaccg aagtgggcga agcgcgccag
      901 ggcactcagc aggacgaagt cattatcgcc gtcggcccgg catttggcct ggcgcaaacc
      961 gtcaatatcg tcggcttacc gcataagagc attctgcgcg aagtcattgc cggtattgaa
     1021 gaagaaggca tcaaggcgcg cgtgattcgc tgctttaaat cttccgacgt ggcgttcgtc
     1081 gccgttgaag gtaaccgcct gagcggatcc ggcatctcca tcggcatcca gtcgaaaggt
     1141 actacggtta tccaccagca ggggctaccg ccgctctcca acctggagct gttcccgcag
     1201 gcaccgctgc tgacgctgga aacctaccgt cagattggta aaaacgccgc ccgctatgcg
     1261 aaacgagaat caccgcagcc ggtccctacg ctcaatgacc agatggcacg cccgaagtac
     1321 caggcaaagt cggccatttt gcatattaaa gagaccaagt acgtcgtgac gggcaaaaac
     1381 ccgcaggaac tgcgcgtggc gctttgaaag ctttcgagta aggatctcca ggcatcaaat
     1441 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1501 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta
     1561 gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa
     1621 tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca
     1681 tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat
     1741 aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct
     1801 caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg
     1861 tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttgaatgc
     1921 accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca
     1981 tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag
     2041 tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag
     2101 ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga
     2161 gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt
     2221 ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg
     2281 taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta
     2341 cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc
     2401 gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca
     2461 aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact
     2521 taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt
     2581 ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca
     2641 gattatattt tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa
     2701 ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc
     2761 ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag
     2821 ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag
     2881 tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca
     2941 cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt
     3001 catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat
     3061 cactatacca attgagatgg gctagtcaat gataattact agtccttttc ccgggtgatc
     3121 tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct
     3181 ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt
     3241 atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact
     3301 cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc
     3361 tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat
     3421 cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac
     3481 attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc
     3541 cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc
     3601 tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag
     3661 ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag
     3721 actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc
     3781 cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     3841 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc
     3901 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     3961 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4021 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4081 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     4141 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     4201 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     4261 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     4321 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     4381 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     4441 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     4501 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     4561 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     4621 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     4681 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     4741 tcagatcatg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     4801 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattcc
//
