LOCUS       sequence-233787-        4244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector into the ROCI-3 site of Rhodococcus opacus PD630 
            containing constitutive EYFP and hygromycin B resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  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 4244)
  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: pDD132
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      51..78
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(106..123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(240..785)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(320..339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1409..1436)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1452..1523)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(1561..2271,2272..2274,2275..2277))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1603..1624)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1952..1971
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2211..2232
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             2284..2295
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(2355..2389)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     primer_bind     complement(2410..2428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gagataggtg ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
       61 cgggtgggcc tttctgcgtt tatacctagg gatatattcc gcttcctcgc tcactgactc
      121 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
      181 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
      241 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
      301 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
      361 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcggttgtct
      421 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
      481 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      541 cccggaaaga catgcaaaag caccactgac agcagccact ggtaattgat ttagaggagt
      601 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
      661 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
      721 cgccctgcaa gacggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
      781 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
      841 cttcaaatgt agcacctgaa gtcagcccca tacgctcgcc ttgtcgtgga tcttctttcc
      901 ggcggcggtc agtccggcgg cgtggtgttc gccttttccc acgtcgatgc cgcaaaacac
      961 ggcgtaactc gtgtccattc agcagtcctt cgtccggcca gtccggtcgc cgatccagca
     1021 tcgatacccg acacccacgt tacgaggtga cctaccgcag tttccggtgg tcctgtccct
     1081 atcagcggtc catcgacgcc accaggttcg gcgacaccac ccccggatca ttcgagagac
     1141 aggggcaaag aagtcatacc gaagatggcg accgcagtcc cgcgaaacgg gaccacaaag
     1201 acggtaacgg caaaccggcc tgggaacccg cgggtttcgc caccctcgaa cgggtcctgg
     1261 cgacgagctg accgcgtcgc ggcgcccgat aagagaccac tcgagctccg accctgattc
     1321 tgtgcgatag tgagttatgc aaccaaatga cgtggctttg ttgaataaat cgaacttttg
     1381 ctgagttgaa ggatcagctc gatataaacg cagaaaggcc cacccgaagg tgagccagtg
     1441 tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     1501 gactgagcct ttcgttttat ttgatgcctg gttatcatga gaccggtctc aattcgcagc
     1561 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     1621 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactggtagc tcaggtagtg
     1681 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     1741 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     1801 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     1861 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     1921 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     1981 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgcagctt
     2041 catgtggtcg gggtagcggg cgaagcattg caggccgtag ccgaaggtgg tcacgagggt
     2101 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     2161 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     2221 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatcta
     2281 gtatttctcc tctttctcta gtatgtgtga aattgttatc cgcttgatct aagcaaacct
     2341 taacagcaga gaaggctagc acaataccta ggactgagct agctgccaac tcgaggtgaa
     2401 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2461 cttagacgtc gatatctggc gaaaatggtt aatcaagggc actctcgtat tgaccaccct
     2521 ggaggagatg atcgaggatg caaggtgcag agccagcctt cttattcggc cttgaattga
     2581 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcttaatga ccatgggcta
     2641 gcgctttagc taattaattg gggaccctag aggtcccctt ttttattttg cggccgccgg
     2701 gggccggggg cggcgccggg cggcccgggg cgtcaggcgc cgggggcggt gtccggcggc
     2761 ccccagagga actgcgccag ttcctccgga tcggtgaagc cggagagatc cagcggggtc
     2821 tcctcgaaca cctcgaagtc gtgcaggaag gtgaaggcga gcagttcgcg ggcgaagtcc
     2881 tcggtccgct tccactgcgc cccgtcgagc agcgcggcca ggatctcgcg gtcgccccgg
     2941 aaggcgttga gatgcagttg caccaggctg tagcgggagt ctcccgcata gacgtcggtg
     3001 aagtcgacga tcccggtgac ctcggtcgcg gccaggtcca cgaagatgtt ggtcccgtgc
     3061 aggtcgccgt ggacgaaccg gggttcgcgg ccggccagca gcgtgtccac gtccggcagc
     3121 cagtcctcca ggcggtccag cagccggggc gagaggtagc cccacccgcg gtggtcctcg
     3181 acggtcgccg cgcggcgttc ccgcagcagt tccgggaaga cctcggaatg gggggtgagc
     3241 acggtgttcc cggtcagcgg caccctgtgc agccggccga gcacccggcc gagttcgcgg
     3301 gccagggcga gcagcgcgtt ccggtcggtc gtgccgtcca tcgcggaccg ccaggtggtg
     3361 ccggtcatcc ggctcatcac caggtagggc cacggccagg ctccggtgcc gggccgcagc
     3421 tcgccgcggc cgaggaggcg gggcaccggc accggggcgt ccgccaggac cgcgtacgcc
     3481 tccgactccg acgcgaggct ctccggaccg caccagtgct cgccgaacag cttgatcacc
     3541 gggccgggct cgccgaccag tacggggttg gtgctctcgc cgggcacccg cagcaccggc
     3601 ggcaccggca gcccgagctc ctccagggct cggcgggcca gcggctccca gaattcctgg
     3661 tcgttccgca ggctcgcgta ggaatcatcc gaatcaatac ggtcgagaag taacagggat
     3721 tcttgtgtca cagcggacct ctattcttgg gtcggatagt gtcgagaact gcgcggtcat
     3781 ggtattcggg acgtttgagt gccgagcaag cggctcgcat cactcggtgg cttcccggtg
     3841 cccgcctgtg tgctgacatg ctttccagtg ctcgagtttg tcgccaaaca tcgagccgga
     3901 gatgaagaac agatcatgta attcaatctc ttcgccgaga cgggtgacga ggcgattgac
     3961 ctcgtcacga ggtacttcgt caacgagttt cgatgagctg tgatagtgct gccaggtgct
     4021 gcgcatctcg tcgagcccat acgtcacttc ggtcgggtcg ttcagcccgg tccttgcagt
     4081 tgcccacaag cagtggttct ccacgatcca cgctcagccg ccgacgccgc cgtgcagcgt
     4141 tggcatacgt gcttcgtgca gtagctgttc gtccgtgtct gccggccact cggtggtgcc
     4201 gccgacgcgg gaccattgcg cgatcaggca tgagaaggtg ctct
//
