LOCUS       sequence-207206-        5269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive promoter expressing araE on RK2 origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5269)
  AUTHORS   Cook TB, Rand JM, Nurani W, Courtney DK, Liu SA, Pfleger BF
  TITLE     Genetic tools for reliable gene expression and recombineering in 
            Pseudomonas putida.
  JOURNAL   J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 
            10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5.
  PUBMED    29299733
REFERENCE   2  (bases 1 to 5269)
  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 Ind 
            Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. 
            doi: 10.1007/s10295-017-2001-5."
COMMENT     Sequence Label: pRK2-AraE
FEATURES             Location/Qualifiers
     source          1..5269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1606..1640)
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     promoter        join(1893..1898,1899..1915,1916..1921)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2110..2643
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     terminator      2915..3009
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3121..3652
                     /label=oriV
                     /note="incP origin of replication"
     CDS             4045..5193
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 agactatcgg ctgtagccta ggttgagaag agaaaagaaa accgccgatc ctgtccaccg
       61 cattactgca aggtagtgga caagaccggc ggtcttaagt tttttggctg aagaattctt
      121 atcagacgcc gatatttctc aacttctcgc ctgccatcag tttgcgttcg atatgttcca
      181 gcgtgacatt tttggtttcc ggaatgagcc agaaagtaat gcccacaaac gcaatgttca
      241 gcgcagtgta gagccagaac gtaccggcag cgccaatgct atcaagcagt gtcaggaagg
      301 tcgcgccgat aatcatattc gacacccagt tcgtggtggt cgaacaggta ataccgaaat
      361 cgcggcattt cagcggctga atttcagagc acaggatcca caccactggc gcggcgctca
      421 tcgcataacc ggcaatacac atcatcgtca tgccaacaga gagccaggac aagccactgg
      481 aagccgtacc gttatcaaac tgcatcaggc aatagcccag caccagagtg cctaacgcca
      541 tcacgctgaa accaattttc agagccggtt tacgccctgc tttatctacc gtaaacaccg
      601 caataaaggt ggcgaacata aaggtcagcc ctacgaccag agtcgcaatc atctgttgtt
      661 ctgtggtcgt aaagcccgcc attttgaaga tacgcggcgc gtagtacatg atgatgttca
      721 taccggtaaa ctgctgcatc gcctgcaaca acataccgag aaacacagca cgacggacgt
      781 tacggttgat cttaaacagt gcccaaccgc cctgttttaa cttcaggctt tcacgaattt
      841 cgttgagttc ttctcgcgct ttttccgacg tatcgcgcag catacgcaat acttcttccg
      901 cctcaatatg acgccccttt tccgccagcc agcgcgggct atttggcagg aagactacca
      961 gaataatcag cagaactgct ggtaaagcaa gaacccccaa cattgcgcgc cagttaccgc
     1021 tataactgaa cgctgtatcg gataaaaacg ccagcacgat gccgagtgtg accatcaact
     1081 ggtacatact gatcatctta ccgcgaacgt tttcacttgc catttcagaa agatacagag
     1141 gagcggtgta agacgcgatc ccgacagcaa tgcccagcac cacacgagcg gcgattaaca
     1201 tctctacgct ggtcgcaaaa gcggacccta tagaaccgag tacaaacagg atggcccccg
     1261 ccatcaggct gtatttacgc cccaggcgga acgacagcca accattaaac agcgcaccaa
     1321 ttgctgcacc gagcatcatg ctactaacca cccattcctg caaacgactg gtcagcacaa
     1381 agtgatcggt aatgaacggc aacgctccgg cgattacgcc gatatcaaga ccaaataaca
     1441 atcctgcgac cgcagcagct accgaaacaa acatattcat acgccgcgta tcccgcaaag
     1501 aacgtggcgt taaagcagat tccgtattga tagtaaccat ggaattaacc tcctacttga
     1561 ctttatgagt tcgtcaacaa tatctcactc gagataagtc gacgtgctag cacagtccct
     1621 aggactgagc tagctgtaaa tatgtatgct agttcaagat ggaccaaaac gaaaaaaggc
     1681 ccccctttcg ggaggcctct tttctggaat ttggtaccga ggacgtcgga attgccagct
     1741 ggggcgccct ctggtaaggt tgggaagcca tggaagccat cacaaacggc atgatgaacc
     1801 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggacgca
     1861 caccgtggaa acggatgaag gcacgaaccc agttgacata agcctgttcg gttcgtaaac
     1921 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1981 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     2041 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     2101 gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     2161 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
     2221 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac
     2281 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
     2341 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
     2401 aggtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggcgagcac
     2461 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
     2521 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
     2581 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
     2641 taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca
     2701 caacgccgcc aggtggcact tttcggggaa atgtgcgcgc ccgcgttcct gctggcgctg
     2761 ggcctgtttc tggcgctgga cttcccgctg ttccgtcagc agcttttcgc ccacggcctt
     2821 gatgatcgcg gcggccttgg cctgcatatc ccgattcaac ggccccaggg cgtccagaac
     2881 gggcttcagg cgctcccgaa ggtgagctcg cttggactcc tgttgataga tccagtaatg
     2941 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     3001 ggtgagaatc caagcactag tgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3061 ctgcggggga tcaggaccgc tgccggagcg caacccactc actacagcag agccatgtag
     3121 ggccgccggc gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg
     3181 gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct
     3241 cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt
     3301 tacgcgagtt tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga
     3361 cacttgaggg gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag
     3421 agtgctgaca gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa
     3481 tccagcattt gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct
     3541 gcttttaaac caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga
     3601 ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg
     3661 cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc
     3721 agccacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     3781 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     3841 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     3901 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     3961 gccaaggatc tgatggcgca ggggatcaag atcgacggat cgatccgggg aattaattcc
     4021 ggggcaatcc cgcaaggagg gtgaatgaat cggacgtttg accggaaggc atacaggcaa
     4081 gaactgatcg acgcggggtt ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc
     4141 atgcgtgcgc cccgcgaaac cttccagtcc gtcggctcga tagtccagca agctacggcc
     4201 aagatcgagc gcgacagcgt gcaactggct ccccctgccc tgcccgcgcc atcggccgcc
     4261 gtggagcgtt cgcgtcgtct cgaacaggag gcggcaggtt tggcgaagtc gatgaccatc
     4321 gacacgcgag gaactatgac gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa
     4381 caggtcagcg aagccaagca ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa
     4441 atgcagcttt ccttgttcga tattgcgccg tggccggaca cgatgcgagc gatgccaaac
     4501 gacacggccc gctctgccct gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg
     4561 caaaacaagg tcattttcca cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag
     4621 ctgcgggccg acgatgacga actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc
     4681 cctatcggcg agccgatcac cttcacgttc tacgagcttt gccaggacct gggctggtcg
     4741 atcaatggcc ggtattacac gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg
     4801 atgggcttca cgtccgaccg cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc
     4861 cgcgtcctgg accgtggcaa gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc
     4921 gtcgtgctgt ttgctggcga ccactacacg aaattcatat gggagaagta ccgcaagctg
     4981 tcgccgacgg cccgacggat gttcgactat ttcagctcgc accgggagcc gtacccgctc
     5041 aagctggaaa ccttccgcct catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc
     5101 gagcaggtcg gcgaagcctg cgaagagttg cgaggcagcg gcctggtgga acacgcctgg
     5161 gtcaatgatg acctggtgca ttgcaaacgc tagggccttg tggggtcagt tccggctggg
     5221 ggttcagcag ccacctgcat cgccctaggt cccgtaagtg cgctgttcc
//
