LOCUS       sequence-125327-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAD eutE1-20- gfpmut2-ssrA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  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 6154)
  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: CMJ119
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      338..793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(425..444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1348..2007)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1922..1941
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2008..2110)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2502..2519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2636..3181)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2716..2735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3253..3272
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3452..4330)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4357..4641
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4564..4583
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4676..4687
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4758..5468
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4758..5468
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4785..4813)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5408..5428
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5549..5566)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5719..5805
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5897..5924
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5943..6034
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
       61 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      121 ttgctgataa atctggagcc ggtgagcgtg agtgtcgcgg tatcattgca gcactggggc
      181 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      241 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      301 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
      361 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      421 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      481 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      541 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      601 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
      661 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
      721 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
      781 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      841 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      901 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      961 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1021 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1081 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
     1141 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     1201 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     1261 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta
     1321 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     1381 ttgtaattca ttaagcattc tgccgacatg gaagccatca cagacggcat gatgaacctg
     1441 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     1501 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     1561 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     1621 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     1681 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     1741 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     1801 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     1861 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     1921 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     1981 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     2041 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     2101 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     2161 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     2221 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     2281 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     2341 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     2401 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     2461 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     2521 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     2581 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     2641 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     2701 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     2761 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     2821 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     2881 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2941 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     3001 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     3061 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     3121 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     3181 agaagatcat cttattaatc agataaaata tttgctcatg agcccgaagt ggcgagcccg
     3241 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     3301 gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg
     3361 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     3421 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     3481 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     3541 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     3601 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     3661 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     3721 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     3781 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     3841 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     3901 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     3961 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     4021 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     4081 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     4141 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     4201 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     4261 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     4321 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     4381 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     4441 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     4501 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     4561 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     4621 aactctctac tgtttctcca tacccgtttt tttgggctag cgaattcgag ctcttaaaga
     4681 ggagaaaggt catgaatcaa caggatattg aacaggtggt gaaagcggta ctgctgaaaa
     4741 tgaaagacag ctctagaagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
     4801 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
     4861 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
     4921 catggccaac acttgtcact actttcgcgt atggtcttca atgctttgcg agatacccag
     4981 atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa
     5041 gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag
     5101 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
     5161 ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca
     5221 aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg
     5281 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     5341 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     5401 atcacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     5461 tatacaaagt cgacgcagca aacgacgaaa actacgcttt agcagcttaa aagcttggct
     5521 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     5581 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     5641 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtcaccc catgcgagag
     5701 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     5761 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     5821 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     5881 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     5941 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6001 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6061 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6121 gaaagtaaaa gatgctgaag atcagttggg tgca
//
