LOCUS       sequence-476152-        4439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5'Homology psbD reverse in the M2 position used for genome 
            integration at the psbD locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4439)
  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 4439)
  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_023_psbD_5'Hom_rev_Cr
FEATURES             Location/Qualifiers
     source          1..4439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2383..2440
                     /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(2464..2481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2635..3223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2715..2734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3406..3500)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3521..4180)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4095..4114
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4342..4360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4364..4407)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4410..4431
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcgcgc tcacaaagca gttctagtcc catatatata actatatata acccgtttaa
       61 agatttattt aaaaatatgt gtgtaaaaaa tgcttatttt taattttatt ttatataagt
      121 tataatatta aatacacaat gattaaaatt aaataataat aaatttaacg taacgatgag
      181 ttgttttttt attttggaga tacacgcaat gacaattgcg atcggtacat atcaagagaa
      241 acgcacatgg ttcgatgacg ctgatgactg gcttcgtcaa gaccgtttcg tattcgtagg
      301 ttggtcaggt ttattactat tcccttgtgc ttactttgca ttaggtggtt ggttaactgg
      361 tactactttc gttacttcat ggtatacgca tggtttagct acttcttact tagaaggttg
      421 taacttctta acagcagctg tttctacacc tgctaacagt atggctcact ctcttctatt
      481 tgtttggggt ccagaagctc aaggtgattt cactcgttgg tgtcaacttg gtggtttatg
      541 ggcattcgtt gctttacacg gtgcatttgg tttaattggt ttcatgcttc gtcagtttga
      601 aattgctcgt tcagtaaact tacgtccata caacgcaatt gctttctcag caccaattgc
      661 tgtattcgtt tcagtattcc taatttaccc attaggtcaa tcaggttggt tctttgcacc
      721 tagtttcggt gtagctgcta tcttccgttt cattttattc ttccaaggtt tccacaactg
      781 gacacttaac ccattccaca tgatgggtgt tgctggtgtt ttaggtgctg ctttattatg
      841 tgctattcac ggtgctactg ttgaaaacac attattcgaa gacggtgacg gtgctaacac
      901 attccgtgca ttcaacccta cacaggctga agaaacatac tctatggtta ctgctaaccg
      961 tttctggtca caaatcttcg gtgttgcttt ctctaacaaa cgttggcttc acttcttcat
     1021 gttattagtt ccagtaactg gtctttggat gagtgctatt ggtgttgtag gtttagctct
     1081 aaacttacgt gcttacgact tcgtatcaca agagattcgt gctgctgaag accctgaatt
     1141 cgaaacattc tacactaaaa acattcttct taacgaaggt attcgtgctt ggatggctgc
     1201 tcaagaccaa ccacacgaac gtttagtatt ccctgaagaa gtattaccac gtggtaacgc
     1261 tctataatat atttttatat aaattaccaa tactaattag tattggtaat ttatattact
     1321 ttattattta aaagaaaatg cccctttggg gctaaaaatc acatgagtgc ttgagccgta
     1381 tgcgaaaaaa ctcgcatgta cggttcttta ggaggattta aaatattaaa aaataaaaaa
     1441 acaaatccta cctgactaaa ccaggacatt tttcacgtac tctgtcaaaa ggtccaaaca
     1501 caacaacttg gatttggaac cttcacgcag atgctcatga ctttgacagt catacaagtg
     1561 atctagaaga aatttctaga aaagtattca gtgcacactt tggtcaatta ggtatcattt
     1621 tcatttggtt aagtgggtgc gacacgaaga cgtatatatt tttatagttt aaaaagatac
     1681 ttttacactg tagttgaaaa gtataagcac ttttaaaaat caaagcagta taaggcaatt
     1741 atttgcattt tgctttagtt ctttttattt tttcagaaaa acttactttt tagtttcaat
     1801 tatgtctaaa aagacagaag tggtttatca tataaaaaga tttttacaat ccactttttt
     1861 aaaaatcttt tggtttttta ttaaaatcta tactcttaat acaaatggca gatcctaaat
     1921 ccttgtaaga catcccctta agggacggtt ttatcaagtt ccatagccta acagaggtaa
     1981 atccatttta ataaccacaa agcgtttatc gttaaaataa aatactaata tatgtaacgt
     2041 gtaaaaataa tattttaata ttattaagga tactcaaaaa aaatacatta acaagtctaa
     2101 tagaacacaa aatgtattaa atcagctagg tctgaaaaaa ctttaattta tttttaagga
     2161 catctctgcc tctttgggta tagcaattca attgatttgc tctatattta tatccctaaa
     2221 ctagataaac tttgtagaaa ctataaattt tatgcagttt atccgaaatt aaagtcaaac
     2281 gtttttttct ttgagtgaat tctttttttc gatttttatt attccaaacc tgttcgagac
     2341 gagactgaga ccactcatcg cgactagtag cggccgctgc agtccggcaa aaaagggcaa
     2401 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc
     2461 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2521 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2581 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2641 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2701 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2761 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2821 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2881 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2941 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3001 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3061 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3241 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3361 ctaaagtata tatgagtaaa cttggtctga cagctcgagg cttggattct caccaataaa
     3421 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     3481 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     3541 cgtcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     3601 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3661 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     3721 tgaaactcac ccagggattg gctgatacga aaaacatatt ctcaataaac cctttaggga
     3781 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     3841 ggaaatcgtc gtggtattca ctccagagtg atgaaaacgt ttcagtttgc tcatggaaaa
     3901 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3961 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     4021 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     4081 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     4141 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     4201 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     4261 tggaacctct tacgtgcccg atcaactcga gtgccacctg acgtctaaga aaccattatt
     4321 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
     4381 tccttagctt tcgctaagga tgatttctgg aattcgcggc cgcttctaga gcgatgagt
//
