LOCUS       sequence-471485-        2889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aminoglycoside-3'-phosphotransferase selection gene, codon optimized
            for Chlamydomonas reinhardtii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2889)
  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 2889)
  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_4_035_aphA6-Kan
FEATURES             Location/Qualifiers
     source          1..2889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      291..385
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      568..1156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1057..1076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1310..1327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1351..1408)
                     /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."
     misc_feature    2249..2270
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      2273..2316
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2320..2338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2500..2889,1..270)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2566..2585)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtatcagc caatccctgg
       61 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
      121 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
      181 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
      241 tactgcgacg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
      301 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
      361 ccgggcgttt tttattggtg agaatccaag cctcgagctg tcagaccaag tttactcata
      421 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      481 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1141 gcggagcctg tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1201 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1261 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1321 gagcgaggaa gcctgcataa cgcgaagtaa tcttttcggt tttaaagaaa aagggcaggg
     1381 tggtgacacc ttgccctttt ttgccggact gcagcggccg ctactagtcg cgatgagtgg
     1441 tctcaaagca ttcaattcat caagttttaa aaaataattc cttttgtcag gtctatcatt
     1501 ttttaaatgc tttaaaaata ttttcgcagt ttcctccgat gcatcctctc ttaggcaacg
     1561 ttcaacaaag gatatatcta caaattcatc tgctaaccca gcacgaccaa gatctaaaaa
     1621 ataaatttca ttgaatttat ctataaaaat attactatcc gtgatatcgc catgagaaaa
     1681 aaccaatctt tcttcaacac gagtctcggt taactcattc catagactta ggtaagtttt
     1741 atggtcaccc cataattcag tgtcaaaatc atcttgatct atatcgtcaa ggagttggtt
     1801 atcaataaaa aattttgact cttttaaccg atgatcaatg tttgaaataa atggacaatc
     1861 aataatagca attgaattta acagattgag tgcctcctta tagatagcaa gcaattcttg
     1921 gtctgttaaa aaaagcgctg aaattggttt tgcattgatc gctttagtga tcatgaattc
     1981 aaactgctca tcctgaaaag tcatgatgag ttcaggcacc tttaatttct cagagagcca
     2041 actcaacatt ttcgcttcac gagagacact gtatgttgtc tctgtatata aagtgctaga
     2101 tcgcttaaga aaaaaagttt cattatttcg attaaaagaa taaacatccg atggcgactg
     2161 accaatttta tttggctcta aaacgctgtt tccgataaat tgttgaataa tatttggtaa
     2221 ttccatggtc attcgagacc actcatcgct ctagaagcgg ccgcgaattc cagaaatcat
     2281 ccttagcgaa agctaaggat tttttttatc tgaaattctg cctcgtgata cgcctatttt
     2341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact cgagttgatc
     2401 gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     2461 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     2521 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     2581 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     2641 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     2701 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     2761 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc actctggagt
     2821 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     2881 ggtgaaaac
//
