LOCUS       sequence-122805-        4849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTET HochAld1-20-pduD21-224- FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  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 4849)
  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: CMJ159
FEATURES             Location/Qualifiers
     source          1..4849
                     /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      1437..1508
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1524..1551
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2082..3032)
                     /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(3794..3888)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3919..4713)
                     /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(4030..4049)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4640..4659
                     /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 atggctctgc gcgaggatcg catcgccgag atcgtcgagc gcgtgctcgc
      781 gcgcctcgac tctggcagcg 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 tgcatattaa agaaaccaag tacgtcgtga cgggcaaaaa
     1381 cccgcaggaa ctgcgcgtgg cgctttgaaa gctttcgagt aaggatctcc aggcatcaaa
     1441 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1501 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacct
     1561 agggtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt tgttatcaga
     1621 atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc cagaattgcc
     1681 atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga
     1741 taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta acaagttgtc
     1801 tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct gttctattag
     1861 gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca gctttgaatg
     1921 caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc
     1981 atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta
     2041 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     2101 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     2161 agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     2221 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat
     2281 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     2341 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     2401 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     2461 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     2521 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     2581 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     2641 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     2701 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     2761 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     2821 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     2881 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     2941 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     3001 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     3061 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cccgggtgat
     3121 ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     3181 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     3241 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     3301 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     3361 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     3421 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     3481 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     3541 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     3601 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     3661 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     3721 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt
     3781 ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     3841 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct
     3901 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3961 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4021 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     4081 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4141 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     4201 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4261 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4321 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4381 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4441 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4501 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     4561 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4621 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4681 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     4741 atcagatcat gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     4801 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattcc
//
