LOCUS       Exported                5391 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optimized aTc inducible pTet promoter for Rhodococcus opacus 
            expressing mCherry on pAL5000/pMB1 backbone with kanamycin 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    pDD131
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  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 5391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5391
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(167..961)
                     /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
                     SHLAPVEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(278..297)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     888..907
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(2754..3342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2834..2853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3525..3543
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3555..3573
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3587..4297
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3732..3750)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4000..4019
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4400..4427
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
       61 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
      121 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
      181 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
      241 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
      301 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
      361 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
      421 cgtcgggcat ccgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
      481 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
      541 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
      601 gcattgcatc agccatgatg gatactttct cgacaggagc aaggtgagat gacaggagat
      661 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
      721 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
      781 ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
      841 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
      901 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
      961 tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc
     1021 ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg
     1081 ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc
     1141 atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgtctgac attcatccgg
     1201 ggtcagcacc gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg
     1261 cgccctgagt gcttgcggca gcgtgaagct agcgtctacc aggactttta cctgtccgac
     1321 ccgttgcaac ggaacccccc acggaacccc cgcgacaccc gctccccaat tgcgttagaa
     1381 cagcggtgga ttgtcggctt cgttgtgggc cttttgagcc gcttcctgtt ctgccgcacg
     1441 ctctttcctc gcccgatagc cgagtcgctt aacggtgtcc agatgcagcc cgaaatgttt
     1501 ggccgtttgc ggccaagagt ggccctcgtc gtcgtgatag gcgcggatgc gttcgcggcg
     1561 tgcagcctgc tcggcgagcc actcgctgcg ttcctgcgcc acgagccgga cgacgtggcg
     1621 ttcggatagt ccggtgattc gagcgccttc ggcggcggtc acgcgccgct ttttgcggac
     1681 agtcggctgc cggttgtagc cgtcgctgta gccgtcgctc atagcaatgc ctccatggct
     1741 gacgcggact ttgcgcgccg cgcaactgtg ctcgccgccg tgcgcgctgc tgcgcccttc
     1801 cgcgagatgg ccgactggcg cgcactgagt gtggcctcgt agaccacgat cccgtccgcc
     1861 caaatgcgcg acttggttgt gatccaacgc caaatgctgt tggcgatggc gcggacctcg
     1921 ctgtccggta gcggtccggg acacacgtcg ttgcacggga actcggcgtt tcgcgcgtgg
     1981 cactcggcat agatcgcgcg gccgagtccg tccacgttcc gggtcggcag gtagatccgc
     2041 atgagggcgg gacgataggc ccacaacctg acggaatcga acagtgcgca attccgccct
     2101 agcggcgtcg gagccgcttt gtacgtggtc tgctgacgcc agcgcggcgg tggcatgttc
     2161 gcgccgagct cggcctcgat gtggctgagt gtgtagagat ctgagtggag ccattccgtt
     2221 tcccaggcga tgtggccggg gtttttggtc atgaggcctg agtaactgcg gtcgccgtcc
     2281 acggcgcgcc gaaggccttc ggcgcacgcc gccatgtatg cgagcggctt acgccgcgcg
     2341 tattcggtgc gtggaacagg ggcgttgagt gcccacactg cgtgtgcgtg gccgttggcg
     2401 cgattgccca cgatcgcgtt gggcagcgga tgggaccccc gggcgctgag cgctcggagc
     2461 gctgcgtctg gatggtctac gtccacgacc agcaggtttg ccagcgctgt tgggttcgcc
     2521 tcgatgtacc ggcggcctag ggccgacgcg cggctttggc ggtagatccc ctcgagcaga
     2581 tcgtcgcttg ccagcggcca gtacggcagc cagagctgct caaattcgtc ggcgacgtgg
     2641 ctcacgcttg gtagtagacc acgattaatc accggtgtat ggtccgacac gagctccaag
     2701 tcagatattt cgcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccctcggg aagcgtggcg
     2941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3181 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3361 tctacggggt ctgacgctca tggaacaata aacgaaaggc tcagtcgaaa gactgggcct
     3421 ttcgtttatc tgttgtttga aaaaaaaaag ccccgcgatt gcggggcttt tttttttcat
     3481 atgtctgacc agggaaaata gccctctgac ctggggattt gacttcccta tcagtgatag
     3541 agataatctg ggagtcccta tcagtgatag agaaggcggt atcgatatgg tgagcaaggg
     3601 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     3661 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3721 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3781 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3841 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3901 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3961 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     4021 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     4081 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     4141 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     4201 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     4261 ccactccacc ggcggcatgg acgagctgta caagtaacca tactctggcc atttttcttt
     4321 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg caacgcgtgc
     4381 ggccgctgat tagctaagca gaaggccatc ctgacggatg gcctttttct agaaaaaaaa
     4441 aagcgccgca actgcggcgc tttttttttt tttaattaag tgatagattg cgatgcgtgt
     4501 gctcggttga acctctctgt cagatcagga ggacttcgca tgtcgcggtt ggataagtcc
     4561 aaggtcatca actcggcgct ggagctgctg aacgaggtcg gtatcgaggg actgaccacg
     4621 cgcaagttgg cccaaaagct cggcgtggaa caaccgaccc tctactggca cgtgaagaac
     4681 aagcgcgcgc tgctggatgc gttggccatc gagatgctgg atcgccatca cacgcacttt
     4741 tgccccctgg agggcgagag ctggcaggat tttctgcgga acaacgcgaa gtcgttccgg
     4801 tgcgccctcc tgagccatcg ggatggcgcc aaagtccact tgggcacccg cccgaccgag
     4861 aagcaatacg agaccctgga gaaccagctg gccttcctct gtcaacaggg tttttcgttg
     4921 gagaacgccc tgtatgccct gtccgccgtg ggacacttca ccttgggctg cgtgttggag
     4981 gatcaggaac atcaggtggc gaaagaggaa cgggagaccc ccacgacgga ttcgatgccc
     5041 ccgttgttgc gccaggcgat cgaactgttt gaccaccagg gcgccgaacc cgcctttttg
     5101 ttcgggctgg agctgatcat ctgcgggctg gaaaagcaac tgaagtgcga aagcggttcg
     5161 tgataacgcc gaaaaccccg cttcggcggg gttttgccgc ggtctgcgtt gtcgggaaga
     5221 tttaaaataa aaaaggggac ctctagggtc cccaattaaa tagttaacac tagtattaaa
     5281 aaaaaaagcc ccgcgattgc ggggcttttt ttttttcgac ggcgagggtt ccgaccgctg
     5341 caactcccgg tgcaaccttg tcccggtcta ttctcttcac tgcaccagct c
//
