LOCUS       sequence-127776-        4972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET pduP1-18-mCherry-ssrA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4972)
  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 4972)
  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: CMJ205
FEATURES             Location/Qualifiers
     source          1..4972
                     /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             785..1489
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(927..945)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1195..1214
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1560..1631
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1647..1674
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2205..3155)
                     /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(3917..4011)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4042..4836)
                     /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(4153..4172)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4763..4782
                     /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 atgaatactt ctgaactcga aaccctgatt cgcaccattc ttagcgagca
      781 attagtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
      841 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      901 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
      961 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1021 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1081 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctccagga
     1141 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1201 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     1261 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     1321 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     1381 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1441 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg tcgacgcagc
     1501 aaacgacgaa aactacgctt tagcagctta aaagctttcg agtaaggatc tccaggcatc
     1561 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1621 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     1681 cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta gtttgttatc
     1741 agaatcgcag atccggcttc agccggtttg ccggctgaaa gcgctatttc ttccagaatt
     1801 gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc agctttgatt
     1861 cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt
     1921 gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat
     1981 taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga
     2041 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     2101 tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg
     2161 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     2221 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     2281 ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     2341 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct
     2401 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     2461 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     2521 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     2581 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     2641 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     2701 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     2761 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     2821 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     2881 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     2941 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     3001 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     3061 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     3121 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     3181 taatcactat accaattgag atgggctagt caatgataat tactagtcct tttcccgggt
     3241 gatctgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     3301 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     3361 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     3421 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     3481 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     3541 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     3601 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     3661 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     3721 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     3781 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     3841 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     3901 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     3961 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     4021 tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     4081 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     4141 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     4201 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     4261 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     4321 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     4381 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     4441 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     4501 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     4561 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     4621 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     4681 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     4741 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     4801 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     4861 ttgatcagat catgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     4921 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt cc
//
