LOCUS       sequence-236072-        4792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The pLPD03031 promoter from Rhodococcus opacus PD630 expressing GFP+
            on the pAL5000/pMB1 backbone with kanamycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4792)
  AUTHORS   DeLorenzo DM, Henson WR, Moon TS
  TITLE     Development of Chemical and Metabolite Sensors for Rhodococcus 
            opacus PD630.
  JOURNAL   ACS Synth Biol. 2017 Oct 20;6(10):1973-1978. doi: 
            10.1021/acssynbio.7b00192. Epub 2017 Aug 10.
  PUBMED    28745867
REFERENCE   2  (bases 1 to 4792)
  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.7b00192"; journalName: "ACS Synth Biol"; date: 
            "2017-10-20- 20"; volume: "6"; issue: "10"; pages: "1973-1978"
COMMENT     Sequence Label: pDD87
FEATURES             Location/Qualifiers
     source          1..4792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..47
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             join(358..360,361..363,364..1077)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNMEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(391..419)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1014..1034
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1102..1119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1273..1861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1353..1372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2145..2939)
                     /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(2256..2275)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2866..2885
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 actagttgat tagctaagca gaaggccatc ctgacggatg gcctttttgc gtttaatact
       61 gtttaaactc tagaaatatt ggatcgtcgg caccgtcacg gccgtgggag gcggcacgat
      121 ccgcgacgtg atgatcggcc gcatccccac ggtgctgcgc agtgagctct acgccatccc
      181 ggcgttgatc tgtgcgttcg ctcggacgct cttcgacacg cctaggaaac atgcaggaaa
      241 cctcacatcg gcccggctga aaagtgagga tcagttgacg ttgcggtcac cgaccgcagc
      301 accgccgcaa caccggtttt ctacctttgg gtctcccaca caacgaggag gcccgggatg
      361 gctagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      421 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc tacatacgga
      481 aagcttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      541 gtcactactt tgacctatgg tgttcaatgc ttttcccgtt atccggatca tatgaaacgg
      601 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
      661 aaagatgacg ggaactacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
      721 aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct cggacacaaa
      781 ctcgagtaca actataactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
      841 atcaaagcta acttcaaaat tcgccacaac atggaagatg gctccgttca actagcagac
      901 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      961 ctgtcgacac aatctgccct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     1021 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaataatga
     1081 attcgagctg gatccagctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1141 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1201 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1441 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1681 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tggggtgggc
     1981 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     2041 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     2101 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     2161 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     2221 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     2281 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     2341 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     2401 tcgggcatcc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     2461 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     2521 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     2581 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     2641 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2701 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgg
     2761 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     2821 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     2881 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     2941 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     3001 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     3061 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     3121 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgtctgacat tcatccgggg
     3181 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
     3241 ccctgagtgc ttgcggcagc gtgaagctag cgtctaccag gacttttacc tgtccgaccc
     3301 gttgcaacgg aaccccccac ggaacccccg cgacacccgc tccccaattg cgttagaaca
     3361 gcggtggatt gtcggcttcg ttgtgggcct tttgagccgc ttcctgttct gccgcacgct
     3421 ctttcctcgc ccgatagccg agtcgcttaa cggtgtccag atgcagcccg aaatgtttgg
     3481 ccgtttgcgg ccaagagtgg ccctcgtcgt cgtgataggc gcggatgcgt tcgcggcgtg
     3541 cagcctgctc ggcgagccac tcgctgcgtt cctgcgccac gagccggacg acgtggcgtt
     3601 cggatagtcc ggtgattcga gcgccttcgg cggcggtcac gcgccgcttt ttgcggacag
     3661 tcggctgccg gttgtagccg tcgctgtagc cgtcgctcat agcaatgcct ccatggctga
     3721 cgcggacttt gcgcgccgcg caactgtgct cgccgccgtg cgcgctgctg cgcccttccg
     3781 cgagatggcc gactggcgcg cactgagtgt ggcctcgtag accacgatcc cgtccgccca
     3841 aatgcgcgac ttggttgtga tccaacgcca aatgctgttg gcgatggcgc ggacctcgct
     3901 gtccggtagc ggtccgggac acacgtcgtt gcacgggaac tcggcgtttc gcgcgtggca
     3961 ctcggcatag atcgcgcggc cgagtccgtc cacgttccgg gtcggcaggt agatccgcat
     4021 gagggcggga cgataggccc acaacctgac ggaatcgaac agtgcgcaat tccgccctag
     4081 cggcgtcgga gccgctttgt acgtggtctg ctgacgccag cgcggcggtg gcatgttcgc
     4141 gccgagctcg gcctcgatgt ggctgagtgt gtagagatct gagtggagcc attccgtttc
     4201 ccaggcgatg tggccggggt ttttggtcat gaggcctgag taactgcggt cgccgtccac
     4261 ggcgcgccga aggccttcgg cgcacgccgc catgtatgcg agcggcttac gccgcgcgta
     4321 ttcggtgcgt ggaacagggg cgttgagtgc ccacactgcg tgtgcgtggc cgttggcgcg
     4381 attgcccacg atcgcgttgg gcagcggatg ggacccccgg gcgctgagcg ctcggagcgc
     4441 tgcgtctgga tggtctacgt ccacgaccag caggtttgcc agcgctgttg ggttcgcctc
     4501 gatgtaccgg cggcctaggg ccgacgcgcg gctttggcgg tagatcccct cgagcagatc
     4561 gtcgcttgcc agcggccagt acggcagcca gagctgctca aattcgtcgg cgacgtggct
     4621 cacgcttggt agtagaccac gattaatcac cggtgtatgg tccgacacga gctccaagtc
     4681 agatatttcg ctgaggggcc accccacaac tgcacactcc cccgctctcc cgtcgagccc
     4741 tggtggtgga acaccagcga cagccgagca cccccaacca cctgtaccaa cc
//
