LOCUS       40924_42774        7672 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Chloroplast transformation vector pTCH-RS2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  AUTHORS   Shao M, Blechl A, Thomson JG.
  TITLE     Small Serine Recombination Systems ParA-MRS and CinH-RS2 Perform 
            Precise Excision of Plastid DNA
  JOURNAL   Plant Biotechnol. J. (2017) In press
  PUBMED    28421718
REFERENCE   2  (bases 1 to 7672)
  AUTHORS   Shao M, Blechl A, Thomson JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2017) Crop Improvement and Utilization, USDA, 800 
            Buchanan Street, Albany, CA 94710, USA
REFERENCE   3  (bases 1 to 7672)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Biotechnol. J. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-JAN-2017) Crop Improvement and Utilization, USDA, 800 Buchanan 
            Street, Albany, CA 94710, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..824
                     /note="plastid sequence targeting insertions in the
                     trnI-trnA intergenic region II; similar to INSD accession 
                     Z00044"
     rep_origin      complement(1073..1661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1835..2692)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2693..2797)
                     /label=AmpR promoter
     misc_feature    3021..4438
                     /note="plastid sequence targeting insertions in the
                     trnI-trnA intergenic region I; similar to INSD accession 
                     Z00044"
     regulatory      4470..4586
                     /note="plastid rRNA operon PEP promoter; Prrn"
                     /regulatory_class="promoter"
     regulatory      4591..4607
                     /note="psbA 5' UTR; similar to NC_001879"
                     /regulatory_class="attenuator"
     CDS             4610..5398
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     protein_bind    complement(5445..5478)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_recomb     5517..5622
                     /note="ParA resolution site-MRS; similar to INSD accession 
                     M59825"
     misc_recomb     5634..5752
                     /note="CinH resolution site-RS2; similar to INSD accession 
                     AF213017"
     misc_recomb     5792..5843
                     /note="Bxb1 attP52 recognition site; similar to INSD
                     accession JQ305140"
     misc_recomb     5904..5960
                     /note="phiC31 attP57 recognition site; similar to INSD
                     accession GU56446"
     regulatory      6017..6166
                     /note="rps16; rhct1 mRNA 3' maturation; similar to
                     NC_001879"
                     /regulatory_class="terminator"
     regulatory      6176..6278
                     /note="psbA; similar to NC_001879"
                     /regulatory_class="promoter"
     regulatory      6279..6363
                     /note="psbA 5' UTR; similar to NC_001879"
                     /regulatory_class="attenuator"
     CDS             6378..7046
                     /label=DsRed1
                     /note="wild-type DsRed"
     misc_recomb     7062..7180
                     /note="similar to INSD accession JQ305140; spacer region
                     has been modified to not match the original RS2; engineered
                     to inhibit homologous recombination in chloroplast; CinH 
                     resolution site recognition site - RS2*"
     regulatory      7205..7395
                     /note="psbA 3' UTR; similar to NC_001879"
                     /regulatory_class="terminator"
     misc_feature    7441..7619
                     /note="plastid DNA targeting sequence; similar to inverted 
                     repeat sequence from INSD accession Z00044"
ORIGIN
        1 ctccacttgg ctcgggggga tatagctcag ttggtagagc tccgctcttg caattgggtc
       61 gttgcgatta cgggttggat gtctaattgt ccaggcggta atgatagtat cttgtacctg
      121 aaccggtggc tcactttttc taagtaatgg ggaagaggac cgaaacgtgc cactgaaaga
      181 ctctactgag acaaagatgg gctgtcaaga acgtagagga ggtaggatgg gcagttggtc
      241 agatctagta tggatcgtac atggacggta gttggagtcg gcggctctcc cagggttccc
      301 tcatctgaga tctctgggga agaggatcaa gttggccctt gcgaacagct tgatgcacta
      361 tctcccttca accctttgag cgaaatgcgg caaaagaaaa ggaaggaaaa tccatggacc
      421 gaccccatca tctccacccc gtaggaacta cgagatcacc ccaaggacgc cttcggcatc
      481 caggggtcac ggaccgacca tagaaccctg ttcaataagt ggaacgcatt agctgtccgc
      541 tctcaggttg ggcagtcagg gtcggagaag ggcaatgact cattcttagt tagaatggga
      601 ttccaactca gcaccttttg agtgagattt tgagaagagt tgctctttgg agagcacagt
      661 acgatgaaag ttgtaagctg tgttcggggg ggagttattg tctatcgttg gcctctatgg
      721 tagaatcagt cgggggacct gagaggcggt ggtttaccct gcggcggatg tcagcggttc
      781 gagtccgctt atctccaact cgtgaactta gccgatacaa agcttatcga taccgtctgc
      841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttcgcctt
      901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1501 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2821 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2881 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2941 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3001 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3061 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3121 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3181 cctcttcgct attacgccag gggccttgta cacaccgccc gtcacactat gggagctggc
     3241 catgcccgaa gtcgttacct taaccgcaag gagggggatg ccgaaggcag ggctagtgac
     3301 tggagtgaag tcgtaacaag gtagccgtac tggaaggtgc ggctggatca cctccttttc
     3361 agggagagct aatgcttgtt gggtattttg gtttgacact gcttcacacc cccaaaaaaa
     3421 agaagggagc tacgtctgag ttaaacttgg agatggaagt cttctttcct ttctcgacgg
     3481 tgaagtaaga ccaagctcat gagcttatta tcctaggtcg gaacaagttg ataggacccc
     3541 cttttttacg tccccatgtt ccccccgtgt ggcgacatgg gggcgaaaaa aggaaagaga
     3601 gggatggggt ttctctcgct tttggcatag cgggccccca gtgggaggct cgcacgacgg
     3661 gctattagct cagtggtaga gcgcgcccct gataattgcg tcgttgtgcc tgggctgtga
     3721 gggctctcag ccacatggat agttcaatgt gctcatcggc gcctgaccct gagatgtgga
     3781 tcatccaagg cacattagca tggcgtactc ctcctgttcg aaccggggtt tgaaaccaaa
     3841 ctcctcctca ggaggataga tggggcgatt cgggtgagat ccaatgtaga tccaactttc
     3901 gattcactcg tgggatccgg gcggtccggg ggggaccacc acggctcctc tcttctcgag
     3961 aatccataca tcccttatca gtgtatggac agctatctct cgagcacagg tttagcaatg
     4021 ggaaaataaa atggagcacc taacaacgca tcttcacaga ccaagaacta cgagatcgcc
     4081 cctttcattc tggggtgacg gagggatcgt accattcgag ccgttttttt cttgactcga
     4141 aatgggagca ggtttgaaaa aggatcttag agtgtctagg gttgggccag gagggtctct
     4201 taacgccttc ttttttcttc tcatcggagt tatttcacaa agacttgcca gggtaaggaa
     4261 gaagggggga acaagcacac ttggagagcg cagtacaacg gagagttgta tgctgcgttc
     4321 gggaaggatg aatcgctccc gaaaaggaat ctattgattc tctcccaatt ggttggaccg
     4381 taggtgcgat gatttacttc acgggcgagg tctctggttc aagtccagga tggcccaggg
     4441 ccgcaatgtg agtttttgta gttggatttg ctcccccgcc gtcgttcaat gagaatggat
     4501 aagaggctcg tgggattgac gtgagggggc agggatggct atatttctgg gagcgaactc
     4561 cgggcgaata tgaagcgcat cgatacaagt gagttgtagg gagggaacca tggcagaagc
     4621 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     4681 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     4741 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     4801 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     4861 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     4921 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     4981 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     5041 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     5101 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     5161 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     5221 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     5281 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     5341 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     5401 aatctaagcc gaattctgca gatatccatc acactggcgg ccgcataact tcgtatagca
     5461 tacattatac gaagttatcg tgattagcac atatgtgggc gtgaggctac agcgcgaggc
     5521 gcattaaggt cgggaaaatg cgctaggcgc atttaaattg cgtattgctg taatgcgcca
     5581 tgccggctag actaggccca aatgggtata cccaatttga ccaaggggat cgtcgtaaat
     5641 tataaacttt aaatatcaaa gttacatgtt atatatggtt aaaaatcatt taaatgttac
     5701 atagttttaa gaacttttat attgtaactt tagggtatac cccaaagtaa cgtgctaggg
     5761 ttacgcgtcg catgactacc agggctctca tgggtttgta ccgtacacca ctgagaccgc
     5821 ggtggttgac cagacaaacc acgaagacac aggtcatcac gagatccaat tcctagatct
     5881 gggccccctt gtgtcatgtc ggcgacccta cgcccccaac tgagagaact caaaggttac
     5941 cccagttggg gcactactcc cgaaaaccgc ttctgacgaa ttccccgggg agcccaaggg
     6001 cacgcgtaag cttaccgaaa ttcaattaag gaaattaaat taaggaaata caaaaagggg
     6061 ggtagtcatt tgtatataac tttgtatgac ttttctcttc tatttttttg tatttcctcc
     6121 ctttcctttt ctatttgtat ttttttatca ttgcttccat tgaattccgt gttctgtaga
     6181 gaagtccgta tttttccaat caacttcatt aaaaatttga atagatctac atacaccttg
     6241 gttgacacga gtatataagt catgttatac tgttgaataa caagccttcc attttctatt
     6301 ttgatttgta gaaaactagt gtgcttggga gtccctgatg attaaataaa ccaagatttt
     6361 accggcgcgc catggcctcc tccgagaacg tcatcaagga gttcatgcgc ttcaaggtgc
     6421 gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     6481 cctacgaggg ccacaacacc gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg
     6541 cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg
     6601 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     6661 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggct
     6721 gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc
     6781 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     6841 tgaagggcga gatccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     6901 tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggact
     6961 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca
     7021 ccgagggccg ccaccacctg ttcctggtac cctaagagct ccgtaaatta taaactttaa
     7081 atatcaaagt tacatgttat atatggttaa aaatcattta aatgttacat agttttaaga
     7141 acttttatat tgtaacttta gggtataccc caaagtaacg ccgaggcggc cgctcgagca
     7201 tgcatctaga gcgatcctgg cctagtctat aggaggtttt gaaaagaaag gagcaataat
     7261 cattttcttg ttctatcaag agggtgctat tgctcctttc tttttttctt tttatttatt
     7321 tactagtatt ttacttacat agactttttt gtttacatta tagaaaaaga aggagaggtt
     7381 attttcttgc atttattcat gattgagtat tctattttga ttttgtattt gtttgggctg
     7441 caggtcaact gcccctatcg gaaataggat tgactaccga ttccgaagga actggagtta
     7501 catctctttt ccattcaaga gttcttatgc gtttccacgc ccctttgaga ccccgaaaaa
     7561 tggacaaatt ccttttctta ggaacacata caagattcgt cactacaaaa aggataatgg
     7621 taacctgcgc cagggaaaag aatagaagaa gcatctgact acttcatgca tg
//
