LOCUS       sequence-158002-        3903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis tagged PP3359 under pBAD regulation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3903)
  AUTHORS   Lo TM, Chng SH, Teo WS, Cho HS, Chang MW
  TITLE     A Two-Layer Gene Circuit for Decoupling Cell Growth from Metabolite 
            Production.
  JOURNAL   Cell Syst. 2016 Aug;3(2):133-43. doi: 10.1016/j.cels.2016.07.012.
  PUBMED    27559924
REFERENCE   2  (bases 1 to 3903)
  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"; doi: 
            "10.1016/j.cels.2016.07"; journalName: "Cell Syst"; date: "2016-08";
            volume: "3"; issue: "2"; pages: "133-43"
COMMENT     Sequence Label: pBbE8k-PP3359-6xHis
FEATURES             Location/Qualifiers
     source          1..3903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..885)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        912..1196
                     /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     1119..1138
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1226..1237
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1712..1729
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1747..1818
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1834..1861
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1900..1971
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1987..2014
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2172..2760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2252..2271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2848..2942)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2973..3767)
                     /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(3084..3103)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3694..3713
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
       61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
      121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
      181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
      241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
      301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
      361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
      421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
      481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
      541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
      601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
      661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
      721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
      781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
      841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
      901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
      961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
     1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
     1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
     1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
     1201 gtttttttgg gaattcaaga gatctaaaga ggagaaagga tctatggcta ggtctgcccg
     1261 tagtaccgac gacgcttgcg tggctgctcc tgtgggagaa ggggtgcttg aagacttgat
     1321 cggctacgcc ttgcgacgcg cgcaattgaa gctgtttcag aaccttattg cccggctctc
     1381 ggcccatgac ctgcgcccgg cccaattttc cgccctggcg atcatcgacc agaaccccgg
     1441 gctgatgcag gccgacctgg cgcgtgcgtt ggcaatcgac cccccgcaag tcgtgccaat
     1501 gctgaacaaa ctggaagagc gcgcgctggc cgtgcgcgtg cggtgcaaac cggacaagcg
     1561 ctcgtatggg attttcctga gcaaatcggg cgaggccctg ttgaaggagt tgaagcacat
     1621 cgccgccgac agcgatcacc aggcgacatc caacctctcg gatgacgaaa ggactgaact
     1681 gttgaggtta ttgaagaaaa tctaccggga ccatcatcat catcatcatt aaggatctcc
     1741 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1801 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1861 gtttatagga tccaaactcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag
     1921 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     1981 cacactggct caccttcggg tgggcctttc tgcgtttata cctagggcgt tcggctgcgg
     2041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2221 agtcaaaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     2641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2821 gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     2881 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     2941 tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3001 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3061 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     3121 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3181 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     3241 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     3301 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     3361 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     3421 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     3481 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     3541 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     3601 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3661 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3721 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     3781 catcctgtct cttgatcaga tcatgatccc ctgcgccatc agatccttgg cggcaagaaa
     3841 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     3901 tcc
//
