LOCUS Exported 4336 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Integration vector into the ROCI-2 site of Rhodococcus opacus PD630 containing constitutive EYFP and hygromycin B resistance marker. ACCESSION . VERSION . KEYWORDS pDD108 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4336 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 592..619 /label=T7Te terminator /note="phage T7 early transcription terminator" terminator 635..706 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(744..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)" 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 //