LOCUS       sequence-233111-        9548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible dCas9sth1 and constitutive sgRNA on pAL5000/pMB1
            backbone with gentamicin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9548)
  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 9548)
  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: pDD143
FEATURES             Location/Qualifiers
     source          1..9548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(264..269,270..286,287..292)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             462..995
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            1083..1192
                     /note="incP origin of transfer"
     terminator      3119..3146
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3306..3333
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3857..4445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3937..3956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4753..5631)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5658..5942
                     /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     5865..5884
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      9445..9472
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtaggacagg tgccacttca tccggggtca gcaccaccgg caagcgccgc gacggccgag
       61 gtcttccgat ctcctgaagc cagggcagat ccgtgcacag caccttgccg tagaagaaca
      121 gcaaggccgc caatgcctga cgatgcgtgg agaccgaaac cttgcgctcg ttcgccagcc
      181 aggacagaaa tgcctcgact tcgctgctgc ccaaggttgc cgggtgacgc acaccgtgga
      241 aacggatgaa ggcacgaacc cagttgacat aagcctgttc ggttcgtaaa ctgtaatgca
      301 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
      361 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct atgcctcggg
      421 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      481 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
      541 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
      601 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
      661 tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg
      721 ttgttggtgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
      781 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
      841 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
      901 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
      961 tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct
     1021 tcccggccgc ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt
     1081 agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     1141 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     1201 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     1261 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     1321 ggaaaagcgc tgcttccctg ctgttttgtg ggccgcaaag cgctccgact taactcaggc
     1381 gttagtccca tcacccgctt tatgcgcgca gccacttatt tcagcgctcc gccctgttcc
     1441 aaaggctttc tagccttttg ggcctccagc gcttgtgcag caagcgccac cagctatcct
     1501 tcttgcaggg ctttctacgt gttccgcttc ctttagcagc ccttgcgccc tgagtgcttg
     1561 cggcagcgtg aagctagcgt ctaccaggac ttttacctgt ccgacccgtt gcaacggaac
     1621 cccccacgga acccccgcga cacccgctcc ccaattgcgt tagaacagcg gtggattgtc
     1681 ggcttcgttg tgggcctttt gagccgcttc ctgttctgcc gcacgctctt tcctcgcccg
     1741 atagccgagt cgcttaacgg tgtccagatg cagcccgaaa tgtttggccg tttgcggcca
     1801 agagtggccc tcgtcgtcgt gataggcgcg gatgcgttcg cggcgtgcag cctgctcggc
     1861 gagccactcg ctgcgttcct gcgccacgaa ccggacgacg tggcgttcgg atagtccggt
     1921 gattcgagcg ccttcggcgg cggtcacgcg ccgctttttg cggacagtcg gctgccggtt
     1981 gtagccgtcg ctgtagccgt cgctcatagc aatgcctcca tggctgacgc ggactttgcg
     2041 cgccgcgcaa ctgtgctcgc cgccgtgcgc gctgctgcgc ccttccgcga gatggccgac
     2101 tggcgcgcac tgagtgtggc ctcgtagacc acgatcccgt ccgcccaaat gcgcgacttg
     2161 gttgtgatcc aacgccaaat gctgttggcg atggcgcgga cctcgctgtc cggtagcggt
     2221 ccgggacaca cgtcgttgca cgggaactcg gcgtttcgcg cgtggcactc ggcatagatc
     2281 gcgcggccga gtccgtccac gttccgggtc ggcaggtaga tccgcatgag ggcgggacga
     2341 taggcccaca acctgacgga atcgaacagt gcgcaattcc gccctagcgg cgtcggagcc
     2401 gctttgtacg tggtctgctg acgccagcgc ggcggtggca tgttcgcgcc gagctcggcc
     2461 tcgatgtggc tgagtgtgta gagatctgag tggagccatt ccgtttccca ggcgatgtgg
     2521 ccggggtttt tggtcatgag gcctgagtaa ctgcggtcgc cgtccacggc gcgccgaagg
     2581 ccttcggcgc acgccgccat gtatgcgagc ggcttacgcc gcgcgtattc ggtgcgtgga
     2641 acaggggcgt tgagtgccca cactgcgtgt gcgtggccgt tggcgcgatt gcccacgatc
     2701 gcgttgggca gcggatggga cccccgggcg ctgagcgctc ggagcgctgc gtctggatgg
     2761 tctacgtcca cgaccagcag gtttgccagc gctgttgggt tcgcctcgat gtaccggcgg
     2821 cctagggccg acgcgcggct ttggcggtag atcccctcga gcagatcgtc gcttgccagc
     2881 ggccagtacg gcagccagag ctgctcaaat tcgtcggcga cgtggctcac gcttggtagt
     2941 agaccacgat taatcaccgg tgtatggtcc gacacgagct ccaagtcaga tatttcgctg
     3001 aggggccacc ccacaactgc acactccccc gctctcccgt cgagccctgg tggtggaaca
     3061 ccagcgacag ccgagcaccc ccaaccacct gtaccaacca ctagttgatt agctaagcag
     3121 aaggccatcc tgacggatgg cctttttgcg tttaatactg tttagctcca agtcagatat
     3181 ttcgctgagg ggccacccca caactgcaca ctcccccgct ctcccgtcga gccctggtgg
     3241 tggaacacca gcgacagccg agcaccccca accacctgta ccaaccacta gttgattagc
     3301 taagcagaag gccatcctga cggatggcct ttttgcgttt aatactgttt aaactctaga
     3361 aatattggat cgtcggcacc gtcacggccg tgggaggcgg cacgatccgc gacgtgatga
     3421 tcggccgcat ccccacggtg ctgcgcagtg agctctacgc catcccggcg ttgatctgtg
     3481 cgttcgctgt gcgggctcta acacgtccta gtatggtagg atgagcaaca atgtggtcgg
     3541 ggtagcgggc gtttttgtac tcgaaagaag ctacaaagat aaggcttcat gccgaaatca
     3601 acaccctgtc attttatggc agggtgtttt tttttgtcga cttggggacc ctagaggtcc
     3661 ccttttttat ttttttggat gaagagcatt atcaatggct cttcacagga agcttaaaaa
     3721 aaaaagcccc gcgattgcgg ggcttttttt ttttaatacg gttatccaca gaatcagggg
     3781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4021 ttctcccctc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcatggaaca ataaacgaaa
     4501 ggctcagtcg aaagactggg cctttcgttt atctgttgtt tgaaaaaaaa aagccccgcg
     4561 attgcggggc tttttttttt catatgtctg accagggaaa atagccctca gctccaagtc
     4621 agatatttcg ctgaggggcc accccacaac tgcacactcc cccgctctcc cgtcgagccc
     4681 tggtggtgga acaccagcga cagccgagca cccccaacca cctgtaccaa ccactagttg
     4741 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4801 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4861 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4921 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4981 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     5041 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     5101 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     5161 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     5221 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     5281 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     5341 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     5401 tcctggcggg aacagtaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     5461 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     5521 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     5581 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     5641 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     5701 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     5761 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     5821 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     5881 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     5941 atacagcgga taaagtagca aagagaagga ggttaggaat gtcggacctg gtgctgggcc
     6001 tggccatcgg catcggctcg gtgggcgtgg gcatcctgaa caaggtgacc ggcgagatca
     6061 tccacaagaa ctcgcgcatc ttcccggccg cccaggccga gaacaacctg gtgcgccgca
     6121 ccaaccgcca gggccgccgc ctggcccgcc gcaagaagca ccgccgcgtg cgcctgaacc
     6181 gcctgttcga ggagtcgggc ctgatcaccg acttcaccaa gatctcgatc aacctgaacc
     6241 cgtaccagct gcgcgtgaag ggcctgaccg acgagctgtc gaacgaggag ctgttcatcg
     6301 ccctgaagaa catggtgaag caccgcggca tctcgtacct ggacgacgcc tcggacgacg
     6361 gcaactcgtc ggtgggcgac tacgcccaga tcgtgaagga gaactcgaag cagctggaga
     6421 ccaagacccc gggccagatc cagctggagc gctaccagac ctacggccag ctgcgcggcg
     6481 acttcaccgt ggagaaggac ggcaagaagc accgcctgat caacgtgttc ccgacctcgg
     6541 cctaccgctc ggaggccctg cgcatcctgc agacccagca ggagttcaac ccgcagatca
     6601 ccgacgagtt catcaaccgc tacctggaga tcctgaccgg caagcgcaag tactaccacg
     6661 gcccgggcaa cgagaagtcg cgcaccgact acggccgcta ccgcacctcg ggcgagaccc
     6721 tggacaacat cttcggcatc ctgatcggca agtgcacctt ctacccggac gagttccgcg
     6781 ccgccaaggc ctcgtacacc gcccaggagt tcaacctgct gaacgacctg aacaacctga
     6841 ccgtgccgac cgagaccaag aagctgtcga aggagcagaa gaaccagatc atcaactacg
     6901 tgaagaacga gaaggccatg ggcccggcca agctgttcaa gtacatcgcc aagctgctgt
     6961 cgtgcgacgt ggccgacatc aagggctacc gcatcgacaa gtcgggcaag gccgagatcc
     7021 acaccttcga ggcctaccgc aagatgaaga ccctggagac cctggacatc gagcagatgg
     7081 accgcgagac cctggacaag ctggcctacg tgctgaccct gaacaccgag cgcgagggca
     7141 tccaggaggc cctggagcac gagttcgccg acggctcgtt ctcgcagaag caggtggacg
     7201 agctggtgca gttccgcaag gccaactcgt cgatcttcgg caagggctgg cacaacttct
     7261 cggtgaagct gatgatggag ctgatcccgg agctgtacga gacctcggag gagcagatga
     7321 ccatcctgac ccgcctgggc aagcagaaga ccacctcgtc gtcgaacaag accaagtaca
     7381 tcgacgagaa gctgctgacc gaggagatct acaacccggt ggtggccaag tcggtgcgcc
     7441 aggccatcaa gatcgtgaac gccgccatca aggagtacgg cgacttcgac aacatcgtga
     7501 tcgagatggc ccgcgagacc aacgaggacg acgagaagaa ggccatccag aagatccaga
     7561 aggccaacaa ggacgagaag gacgccgcca tgctgaaggc cgccaaccag tacaacggca
     7621 aggccgagct gccgcactcg gtgttccacg gccacaagca gctggccacc aagatccgcc
     7681 tgtggcacca gcagggcgag cgctgcctgt acaccggcaa gaccatctcg atccacgacc
     7741 tgatcaacaa ctcgaaccag ttcgaggtgg acgccatcct gccgctgtcg atcaccttcg
     7801 acgactcgct ggccaacaag gtgctggtgt acgccaccgc caaccaggag aagggccagc
     7861 gcaccccgta ccaggccctg gactcgatgg acgacgcctg gtcgttccgc gagctgaagg
     7921 ccttcgtgcg cgagtcgaag accctgtcga acaagaagaa ggagtacctg ctgaccgagg
     7981 aggacatctc gaagttcgac gtgcgcaaga agttcatcga gcgcaacctg gtggacaccc
     8041 gctacgcctc gcgcgtggtg ctgaacgccc tgcaggagca cttccgcgcc cacaagatcg
     8101 acaccaaggt gtcggtggtg cgcggccagt tcacctcgca gctgcgccgc cactggggca
     8161 tcgagaagac ccgcgacacc taccaccacc acgccgtgga cgccctgatc atcgccgcct
     8221 cgtcgcagct gaacctgtgg aagaagcaga agaacaccct ggtgtcgtac tcggaggacc
     8281 agctgctgga catcgagacc ggcgagctga tctcggacga cgagtacaag gagtcggtgt
     8341 tcaaggcccc gtaccagcac ttcgtggaca ccctgaagtc gaaggagttc gaggactcga
     8401 tcctgttctc gtaccaggtg gactcgaagt tcaaccgcaa gatctcggac gccaccatct
     8461 acgccacccg ccaggccaag gtgggcaagg acaaggccga cgagacctac gtgctgggca
     8521 agatcaagga catctacacc caggacggct acgacgcctt catgaagatc tacaagaagg
     8581 acaagtcgaa gttcctgatg taccgccacg acccgcagac cttcgagaag gtgatcgagc
     8641 cgatcctgga gaactacccg aacaagcaga tcaacgacaa gggcaaggag gtgccgtgca
     8701 acccgttcct gaagtacaag gaggagcacg gctacatccg caagtactcg aagaagggca
     8761 acggcccgga gatcaagtcg ctgaagtact acgactcgaa gctgggcaac cacatcgaca
     8821 tcaccccgaa ggactcgaac aacaaggtgg tgctgcagtc ggtgtcgccg tggcgcgccg
     8881 acgtgtactt caacaagacc accggcaagt acgagatcct gggcctgaag tacgccgacc
     8941 tgcagttcga caagggcacc ggcacctaca agatctcgca ggagaagtac aacgacatca
     9001 agaagaagga gggcgtggac tcggactcgg agttcaagtt caccctgtac aagaacgacc
     9061 tgctgctggt gaaggacacc gagaccaagg agcagcagct gttccgcttc ctgtcgcgca
     9121 ccatgccgaa gcagaagcac tacgtggagc tgaagccgta cgacaagcag aagttcgagg
     9181 gcggcgaggc cctgatcaag gtgctgggca acgtggccaa ctcgggccag tgcaagaagg
     9241 gcctgggcaa gtcgaacatc tcgatctaca aggtgcgcac cgacgtgctg ggcaaccagc
     9301 acatcatcaa gaacgagggc gacaagccga agctggactt ctaaatactc tggccatttt
     9361 tctttaccgc ctttgagtga gctgataccg ctcgccgcag ccaaacgacc gagcgcaacg
     9421 cgtgcggccg ctgattagct aagcagaagg ccatcctgac ggatggcctt tttctagaaa
     9481 aaaaaagcgc cgcaactgcg gcgctttttt tttttttaat taagtgatag attgcgatgc
     9541 gtgtgctc
//
