LOCUS       sequence-247738-        4336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector into the ROCI-2 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 4336)
  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 4336)
  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: pDD108
FEATURES             Location/Qualifiers
     source          1..4336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(592..619)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(635..706)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(744..1454,1455..1457,1458..1460))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(786..807)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1135..1154
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1394..1415
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             1467..1478
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(1538..1572)
                     /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(1593..1611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3511..3538
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3566..3583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3700..4245)
                     /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(3780..3799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagatacgt gtgctgatcc aacgcctgcg cactcatcac gaaccccttc cattcgaaag
       61 aatcgaaaca cttttcgaat ctttcgaata gctggttcaa tcaccgccgc ttacatggcg
      121 agatcgacca cgtcccacct gccgagtacg agaccgccta ttggtcgaat cacacggcgg
      181 cactacggtg agacaccggt cctcgctgag gccggaacca gataaccgag cctccgccga
      241 actcggggca ggtgtcaccg ccccggtcct cgagagcaag ctcgacggtg tcagcgaaat
      301 gcgtgaacac ctcagccgcg aactggccca gcagcccacc cgccgcatcc ctatccggct
      361 cagcgacgaa tgcacccgcg ccaacaccgc cgcccgcaca gccagccaga acctcgaaac
      421 ctgaatccgc gaatgtgtgc ggtcgccgcc gcagtgctcg ggcattccgg ccgaaccgtt
      481 gtgcctgtct caatccctga ttctgtgcga tagtgagtta tgcaaccaaa tgacgtggct
      541 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag ctcgatataa acgcagaaag
      601 gcccacccga aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga
      661 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggttatca
      721 tgagaccggt ctcaattcgc agccttgtac agctcgtcca tgccgagagt gatcccggcg
      781 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
      841 gcggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
      901 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
      961 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1021 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1081 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1141 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1201 gacttgaaga agtcgtgcag cttcatgtgg tcggggtagc gggcgaagca ttgcaggccg
     1261 tagccgaagg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1321 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1381 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1441 tcctcgccct tgctcaccat ctagtatttc tcctctttct ctagtatgtg tgaaattgtt
     1501 atccgcttga tctaagcaaa ccttaacagc agagaaggct agcacaatac ctaggactga
     1561 gctagctgtc aactcgaggt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     1621 taatgtcatg ataataatgg tttcttagac gtcgatatct ggcgaaaatg gttaatcaag
     1681 ggcactctcg tattgaccac cctggaggag atgatcgagg atgcaaggtg cagagccagc
     1741 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     1801 tgggtcttaa tgaccatggg ctagcgcttt agctaattaa ttggggaccc tagaggtccc
     1861 cttttttatt ttgcggccgc cgggggccgg gggcggcgcc gggcggcccg gggcgtcagg
     1921 cgccgggggc ggtgtccggc ggcccccaga ggaactgcgc cagttcctcc ggatcggtga
     1981 agccggagag atccagcggg gtctcctcga acacctcgaa gtcgtgcagg aaggtgaagg
     2041 cgagcagttc gcgggcgaag tcctcggtcc gcttccactg cgccccgtcg agcagcgcgg
     2101 ccaggatctc gcggtcgccc cggaaggcgt tgagatgcag ttgcaccagg ctgtagcggg
     2161 agtctcccgc atagacgtcg gtgaagtcga cgatcccggt gacctcggtc gcggccaggt
     2221 ccacgaagat gttggtcccg tgcaggtcgc cgtggacgaa ccggggttcg cggccggcca
     2281 gcagcgtgtc cacgtccggc agccagtcct ccaggcggtc cagcagccgg ggcgagaggt
     2341 agccccaccc gcggtggtcc tcgacggtcg ccgcgcggcg ttcccgcagc agttccggga
     2401 agacctcgga atggggggtg agcacggtgt tcccggtcag cggcaccctg tgcagccggc
     2461 cgagcacccg gccgagttcg cgggccaggg cgagcagcgc gttccggtcg gtcgtgccgt
     2521 ccatcgcgga ccgccaggtg gtgccggtca tccggctcat caccaggtag ggccacggcc
     2581 aggctccggt gccgggccgc agctcgccgc ggccgaggag gcggggcacc ggcaccgggg
     2641 cgtccgccag gaccgcgtac gcctccgact ccgacgcgag gctctccgga ccgcaccagt
     2701 gctcgccgaa cagcttgatc accgggccgg gctcgccgac cagtacgggg ttggtgctct
     2761 cgccgggcac ccgcagcacc ggcggcaccg gcagcccgag ctcctccagg gctcggcggg
     2821 ccagcggctc ccagaattcc tggtcgttcc gcaggctcgc gtaggaatca tccgaatcaa
     2881 tacggtcgag aagtaacagg gattcttgtg tcacagcgga cctctattcg gtactgcgcg
     2941 agagggtaaa cggctgccac ccgtccctgt cgccgtacgc gggcatcagc gacgctcccc
     3001 aaacgtcgac tgccactgcc actgccactg ccactgccac tgccactgag caacgaactc
     3061 gacgagctcg ggcagcgcca gcgctgcaag cccagcagaa ctcatgcgcg gggcggtcga
     3121 ggaatacatc ttcgcctgaa acaccccccg acgctcgcgg agcaacctgt cgaaacacca
     3181 cctcggattc cgtcgcacaa tcgaaaggga ggagaaggcg gaaggacctg ctggaggtga
     3241 ccatgcgtcg ggaggaactg gcaccggtgc ccacgccggt ccggtggtgg cggctcggcc
     3301 catggagccg caaccacctg atgcgcccca gcgaccgcta cgaggccacc atcatcctca
     3361 tcatcctgct caccgcggcc gtggtcctgc tgctgatccc cgttgcggcc gcgttcggca
     3421 cctcgacgca gacccggttg gaacagcaaa cccacacact gagataggtg ctgttgtttg
     3481 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     3541 tatacctagg gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     3601 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     3661 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     3721 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     3781 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
     3841 gtttaccggt gtcattccgc tgttatggcc gcggttgtct cattccacgc ctgacactca
     3901 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
     3961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
     4021 caccactgac agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
     4081 ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct
     4141 cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa gacggttttt
     4201 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
     4261 aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
     4321 gtcagcccca tacgct
//
