LOCUS       sequence-476150-        4224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5'Homology rbcL reverse in the M2 position used for genome 
            integration at the rbcL locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  AUTHORS   Inckemann R, Chotel T, Brinkmann CK, Burgis M, Andreas L, Baumann J,
            Sharma P, Klose M, Barrett J, Ries F, Paczia N, Glatter T, Willmund 
            F, Mackinder LCM, Erb TJ
  TITLE     Advancing chloroplast synthetic biology through high-throughput 
            plastome engineering of Chlamydomonas reinhardtii
  JOURNAL   bioRxiv 2024.05.08.593163
REFERENCE   2  (bases 1 to 4224)
  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"; journalName: "bioRxiv 
            2024.05.08.593163"
COMMENT     Sequence Label: pME_Cp_0_6_022_rbcL_5'Hom_rev_Cr
FEATURES             Location/Qualifiers
     source          1..4224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2168..2225
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2249..2266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2420..3008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2500..2519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3191..3285)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3306..3965)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3880..3899
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4127..4145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4149..4192)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4195..4216
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcgcgc tgtaagtaaa ctgcgtaaga cgaccgacat atacctaaag gccctttcta
       61 tgctcgactg ataagacaag tacataaatt tgctagttta cattattttt tatttctaaa
      121 tatataatat atttaaatgt atttaaaatt tttcaacaat ttttaaatta tatttccgga
      181 cagattattt taggatcgtc aaaagaagtt acatttattt atataaatgg ttccacaaac
      241 agaaactaaa gcaggtgctg gattcaaagc cggtgtaaaa gactaccgtt taacatacta
      301 cacacctgat tacgtagtaa gagatactga tattttagct gcattccgta tgactccaca
      361 accaggtgtt ccacctgaag aatgtggtgc tgctgtagct gctgaatctt caacaggtac
      421 atggactaca gtatggactg acggtttaac aagtcttgac cgttacaaag gtcgttgtta
      481 cgatatcgaa ccagttccgg gtgaagacaa ccaatacatt gcttacgtag cttacccaat
      541 cgacttattc gaagaaggtt cagtaactaa catgttcact tctattgtag gtaacgtatt
      601 cggtttcaaa gctttacgtg ctctacgtct tgaagacctt cgtattccac ctgcttacgt
      661 taaaacattc gtaggtcctc cacacggtat tcaggtagaa cgtgacaaat taaacaaata
      721 tggtcgtggt cttttaggtt gtacaatcaa acctaaatta ggtctttcag ctaaaaacta
      781 cggtcgtgca gtttatgaat gtttacgtgg tggtcttgac tttactaaag acgacgaaaa
      841 cgtaaactca caaccattca tgcgttggcg tgaccgtttc cttttcgttg ctgaagctat
      901 ttacaaagct caagcagaaa caggtgaagt taaaggtcac tacttaaacg ctactgctgg
      961 tacttgtgaa gaaatgatga aacgtgcagt atgtgctaaa gaattaggtg tacctattat
     1021 tatgcacgac tacttaacag gtggtttcac agctaacact tcattagcta tctactgtcg
     1081 tgacaacggt cttcttctac acatccaccg tgctatgcac gcggttattg accgtcaacg
     1141 taaccacggt attcacttcc gtgttcttgc taaagctctt cgtatgtctg gtggtgacca
     1201 ccttcactct ggtactgttg taggtaaact agaaggtgaa cgtgaagtta ctctaggttt
     1261 cgtagactta atgcgtgatg actacgttga aaaagaccgt agccgtggta tttacttcac
     1321 tcaagactgg tgttcaatgc caggtgttat gccagttgct tcaggtggta ttcacgtatg
     1381 gcacatgcca gctttagttg aaatcttcgg tgatgacgca tgtcttcagt tcggtggtgg
     1441 tactctaggt cacccttggg gtaacgctcc aggtgctgca gctaaccgtg tagctcttga
     1501 agcttgtact caagctcgta acgaaggtcg tgaccttgct cgtgaaggtg gcgacgtaat
     1561 tcgttcagct tgtaaatggt caccagaact tgctgctgca tgtgaagttt ggaaagaaat
     1621 taaattcgaa tttgatacta ttgacaaact ttaattttta tttttcatga tgtttatgtg
     1681 aatagcataa acatcgtttt tatttttatg gtgtttaggt taaataccta aacatcattt
     1741 tacattttta aaattaagtt ctaaagttat cttttgttta aatttgcctg tctttataaa
     1801 ttacgatgtg ccagaaaaat aaaatcttag ctttttatta tagaatttat ctttatgtat
     1861 tatattttat aagttataat aaaagaaata gtaacatact aaagcggatg taactcaatc
     1921 ggtagagtgc gatccttcca agttcgaggt tgtgggttcg agtcccatca tccgctaaac
     1981 caatctataa aagttgttga atatgctgaa atgttttcaa agaaaaagcc tagtttttct
     2041 tttacaacaa gcaaagaaca attggcattc tttgattgta agaaaatgcg cttacgtgga
     2101 gttatgtgtt tttactgttc gagacgagac tgagaccact catcgcgact agtagcggcc
     2161 gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
     2221 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2821 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct
     3181 cgaggcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     3241 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     3301 tcaaattacg ccccgccctg ccactcgtcg cagtactgtt gtaattcatt aagcattctg
     3361 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     3421 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     3481 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga tacgaaaaac
     3541 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     3601 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagtgatgaa
     3661 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     3721 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     3781 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     3841 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     3901 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     3961 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     4021 gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcgagtgcc
     4081 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4141 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc
     4201 gcggccgctt ctagagcgat gagt
//
