LOCUS       sequence-122809-        4978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET pduD1-20-mCherry-ssrA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4978)
  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 4978)
  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: CMJ206
FEATURES             Location/Qualifiers
     source          1..4978
                     /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..1495
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MKVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMN
                     FEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGAL
                     KGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYER
                     AEGRHSTGGMDELYK"
     primer_bind     complement(933..951)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1201..1220
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1566..1637
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1653..1680
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2211..3161)
                     /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(3923..4017)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4048..4842)
                     /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(4159..4178)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4769..4788
                     /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 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      841 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
      901 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
      961 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1021 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     1081 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1141 ccaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1201 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1261 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1321 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1381 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1441 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggtcga
     1501 cgcagcaaac gacgaaaact acgctttagc agcttaaaag ctttcgagta aggatctcca
     1561 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1621 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     1681 tttataccta gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt
     1741 gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc
     1801 agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct
     1861 ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa
     1921 caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg
     1981 ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag
     2041 ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     2101 catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     2161 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     2221 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2281 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     2341 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     2401 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     2461 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     2521 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     2581 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     2641 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     2701 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     2761 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     2821 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     2881 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     2941 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     3001 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     3061 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     3121 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     3181 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     3241 ccgggtgatc tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3301 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     3361 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     3421 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     3481 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     3541 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     3601 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     3661 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     3721 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     3781 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     3841 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg
     3901 tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     3961 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     4021 gcgagctctc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     4081 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     4141 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     4201 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     4261 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt
     4321 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     4381 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     4441 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     4501 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     4561 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     4621 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc
     4681 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     4741 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     4801 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     4861 tgtctcttga tcagatcatg atcccctgcg ccatcagatc cttggcggca agaaagccat
     4921 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattcc
//
