LOCUS       sequence-227256-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of Che9c60 and Che9c61 recombinases on 
            pB264/pBR322 backbone with kanamycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  AUTHORS   DeLorenzo DM, Rottinghaus AG, Henson WR, Moon TS
  TITLE     Molecular Toolkit for Gene Expression Control and Genome 
            Modification in Rhodococcus opacus PD630.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):727-738. doi: 
            10.1021/acssynbio.7b00416. Epub 2018 Feb 5.
  PUBMED    29366319
REFERENCE   2  (bases 1 to 6247)
  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: 
            "10.1021/acssynbio.7b00416"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "727-738"
COMMENT     Sequence Label: pDD120
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      127..715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     616..635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2433..3227)
                     /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(2544..2563)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3154..3173
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      3754..3781
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      6113..6159
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
       61 ttcacgaggc agacctcact agttccactg agcgtcagac cccgtagaaa agatcaaagg
      121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      241 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
      301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      481 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
      541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      781 tcctgcgagg aacgtcgggc ggtcgagcgc ggcgtcgtcg cgtgtggagt tgcgcgagcg
      841 ggtggtcgag gtcgagaagg tggtgcatcg gccggccacg acggccattg ctcgtttgaa
      901 caggtctcac agaccatcag ggtgcaacgc acccgcacgg cagggctgtc ccaaggggga
      961 cgggctggta gtggacgtca tcgcagctga tactcatcgc tcgattcctg ccagggcgtg
     1021 atggacagtg ccgaccgagc atcccagctc attggcgata gctctcatgg acatgccagt
     1081 accgcgtagc tcccgtatct tctcgcgccg ttgctgtgcc cgggacaggt aggcctctcg
     1141 cggctctgat gtccaccgga tgatgctggc cgtcgatacg cctgtgcgtt cggcgagctc
     1201 ccgcactgtt atcccgttgc gaggcagacg ctcagtcatg tgtggcctcc tcaattgcca
     1261 gtcgcacctg atccagtttg gtgctgcgtg cacgccgggc ctggccgctg gcgacgccgc
     1321 ctttgcggcc acgggcggat tggatggtcg agagtgtggc ttcgtagacg gcgataccgt
     1381 ctttccagat gcgggatcgc gtggtgatcc atcggtgaat gctggcggcg atggcgcggg
     1441 cttcagaggg tgggagttcc tcggtgtagc tggcgttgag ggcggtgact tcgcggtgga
     1501 gggcgagttc gaggccggcg ctgtcatgag tgggcaggta gttgcgcatg aggccggggc
     1561 gggtggccca ggtgcgagcg gcgtggaaga ttgcgcagtt gcggccgagt cctatggggg
     1621 cgtcacggtg ggccttggtg tgctgccagc gcggggaggg catgtgtgtg ccgagttcgg
     1681 cctcgagctc ggcgaggctg cgcggttcgg tgtggatcca gtgcgtgtcc cagcctgagt
     1741 gtgtggggtt cttggtcatc aggcctgagt agccgatgtc cccttcgacg gcccggcgta
     1801 gcccttcggt aacggcggcg gcgtacgcga ggggcttccg tcgggcgtac tcggtgcggg
     1861 tgaacggctc gcgcagccac cagtgcaggt gcgcatgccc gttgctgctg ttttcgatca
     1921 cggcgttcgg catagggtga ttgcctgccg ctgacagtgc gcgcagggcg gagtcggggt
     1981 ggtcgacgtc gacgacgagc agattgctca gtgtctgcgg gttggcctcg atgtagcgcc
     2041 gctcaagtgc ggcagggcgg cgcattcgat agaccccctc aaggaagtcg tttgttgcca
     2101 aaggccagaa cggtagccac atttgctccc agtccccacc ttcccgcacc ccgtgtgtgt
     2161 ccatgctggt gaccgtcgca tgctctcgaa cgaaacctgg gcatttccct cggtgtgttc
     2221 aaagttaatc gtgaagccaa gtcagatgtt tcggtcttac ataaacgcga agaactccag
     2281 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     2341 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     2401 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     2461 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     2521 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     2581 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     2641 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatccgc
     2701 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     2761 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     2821 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     2881 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     2941 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     3001 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttggag ttcattcagg
     3061 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3121 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3181 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     3241 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     3301 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     3361 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     3421 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     3481 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtgtt ccgcttcctt
     3541 tagcagccct tgcgccctga gtgcttgcgg cagcgtgaag ctagcgtcta ccaggacttt
     3601 tacctgtccg aagctccaag tcagatattt cgctgagggg ccaccccaca actgcacact
     3661 cccccgctct cccgtcgagc cctggtggtg gaacaccagc gacagccgag cacccccaac
     3721 cacctgtacc aaccactagt tgattagcta agcagaaggc catcctgacg gatggccttt
     3781 ttgcgtttaa tactgtttaa actctagaaa tattggatcg tcggcaccgt cacggccgtg
     3841 ggaggcggca cgatccgcga cgtgatgatc ggccgcatcc ccacggtgct gcgcagtgag
     3901 ctctacgcca tcccggcgtt gatctgtgcg ttcgctgtgc gggctctaac acgtcctagt
     3961 atggtaggat gagcaacatt tcgacgccga gagattcgcc gcccgaaatg agcacgatcc
     4021 gcatgcttaa ttaagaagga gatatacata tgagtgtgcc cacacaggac ggaatgcacc
     4081 ggttcgtcga cgaggacgtc taccacgctg accggggctc gctgtcggta tccggcgcga
     4141 agctgctgtt gccgccgtcg tgtcccgcga aattccgctg ggagatggac aacacccgga
     4201 agccgaaaaa ggtctgggac ttcggacatg tcgcgcacaa actggtgctc ggcaagggtg
     4261 ccgagttcga gatcctcgac cccgaggtgc acgggctgaa ggcggacggt acgccgtcgg
     4321 agaagccgac cgcgacgggc atgtggcgca aggccgaggc tgaggctcgc aaacagggca
     4381 aggtgccgat tcacgtcgac ctgttcacga aggcgtacga catggccgaa aaggtgcgtc
     4441 agcacccgac agccggcccg atcttcgcca atcctgacgg cgaggccgag gtcgcgctgt
     4501 actacaccga ccccgagacc ggcgtgcggc tgcgtggccg gatcgactgg ctcactgacg
     4561 atatcgatga ttacaagacg tcgatgaccg cgaacccggc cgagctgaaa accaagttct
     4621 acaagctcgg ctatttcatg caggcggcct ggtacatcga tctactggtc gccctcgggc
     4681 tcgccgagaa cccgcgattc cggttcatca cgcaggagaa agaaccgccc tacgtcgtga
     4741 ctccgatcca gtacgacgac gaggcgatcg aagaggggcg gcgccgcaac cgccaggcga
     4801 tccggctcta cgccgactgc atggaatcgg gcaagtggcc tgactacagc gacgacgtgg
     4861 tcacgatcag cctgccctcg tgggggctgc cgcgaccgca gaccgtcggc gacgtcgtca
     4921 ccgacagcta tatctacgac accgacccgc tcgaagaggc cgacccgatt gaaggggatt
     4981 acatctatgg ctgaaaatgc tgtcaccaag caggattcgc ccaaggcacc cgagacgatc
     5041 tcgcaagtgc tgcaggtgct cgtgccgcag ctggcgcgtg cagtgcccaa gggtatggac
     5101 cccgaccgca tcgcgcggat cgtgcaaacc gagatccgca agtcgcgcaa cgcgaaagct
     5161 gctggcatcg ctaagcagtc cctcgacgac tgcacgcaag agtcatttgc cggtgcgctg
     5221 ctgacctcgg ccgcgctcgg tctcgagccc ggtgtcaacg gcgagtgcta cctcgtgccc
     5281 taccgcgaca cccggcgcgg cgtggtcgag tgccagctga tcatcggcta ccagggcatc
     5341 gtcaaactgt tctggcagca cccgcgcgcc tcccggatcg acgcgcagtg ggtcggcgcg
     5401 aacgacgaat tccattacac catgggcctc aatccgacgc tgaaacacgt gaaggccaag
     5461 ggtgatcggg gtaatccggt ctacttctac gcgatcgtcg aggtgaccgg cgctgagccg
     5521 ctgtgggacg tgttcaccgc cgacgagatc agggaattgc gtcgcggcaa ggtcggatcc
     5581 tcgggcgaca tcaaggaccc gcagcgctgg atggagcgga agaccgcgct caaacaggtg
     5641 ctgaagctgg caccgaagac gacgcggctc gacgcggcga tccgcgccga cgatcgcccg
     5701 ggcaccgacc tgtcacagtc gcaggcgctc gcgctgccgt cgaccgtcaa gccgacggcc
     5761 gactacatcg acggcgagat cgccgagccg cacgaggtcg atacgccacc gaagtcgtcg
     5821 cgcgcacaac gcgcgcagcg cgccaccgcg ccggcgcccg acgtgcagat ggccaatccc
     5881 gatcagctga agcgcctcgg cgagatccag aaggccgaaa agtacaacga cgccgattgg
     5941 ttcaagttcc tcgccgattc ggccggcgtc aaagccacgc gcgccgccga cctcacgttc
     6001 gacgaggcga aggctgtcat cgacatgttc gacgggccca acgcatgagc gccccggcga
     6061 attccgacgc ggtggttgat ctgcaactag cgtacgatcg actgccaggc atcaaataaa
     6121 acgaaaggct cagtcgaaag actgggcctt tcgttttatg ccatcatggc cgcggtgatc
     6181 agctagccac ctgacgtcgg ggggggggga aagccacgtt gtgtctcaaa atctctgatg
     6241 ttacatt
//
