LOCUS       sequence-471482-        3546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aac6-aph2 tobramycin resistance gene, codon optimized for 
            Chlamydomonas reinhardtii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3546)
  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 3546)
  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_022_aac6-aph2-marker
FEATURES             Location/Qualifiers
     source          1..3546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..23)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      618..712
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      895..1483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1384..1403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1637..1654
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1678..1735)
                     /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    3233..3254
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      3257..3300
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(3304..3322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(3484..3546,1..597)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
       61 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
      121 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
      181 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
      241 ttttcatcac tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
      301 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
      361 atgtttttcg tatcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
      421 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
      481 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
      541 ggcagaatgc ttaatgaatt acaacagtac tgcgacgagt ggcagggcgg ggcgtaattt
      601 gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
      661 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcct
      721 cgagctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
      781 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
      841 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
      901 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      961 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1021 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1081 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1141 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1201 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1261 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1321 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1381 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1441 tcgatttttg tgatgctcgt caggggggcg gagcctgtgg aaaaacgcca gcaacgcggc
     1501 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1561 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     1621 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcc tgcataacgc gaagtaatct
     1681 tttcggtttt aaagaaaaag ggcagggtgg tgacaccttg cccttttttg ccggactgca
     1741 gcggccgcta ctagtcgcga tgagtggtct caaagcgtct ttgtatgtac gtttgtaaat
     1801 ttctttacga ccgttttcaa tgaattcttg tttaatgttt ttaataccgt atacaattgt
     1861 ttcaattgga tagtattctt caacaatgtc ttggtattct tttgcttttt caatatcaat
     1921 gttaccatac atacgtaaaa tgtcttcacc aaagttagta ccaatttctt cttcagagtc
     1981 ttctaataag taaataaagt cacaatattc atcaataata cctgagtcac caaaatcaat
     2041 aatacctgtt aaacggttgt taccgtctaa taataaatgg ttacaactga agtcgttgtg
     2101 acataaacat tttttacctt cgaatacagt tgttgcgttt aaacgttcca tgaaagattc
     2161 aatgtaatct ttttcaatat cagttaagtc gttgtaaata gtttcacgta ataaaatata
     2221 ttcttctaat acgttttgtt tgttatcaat agtacattca gaaatatcag tgtagtctaa
     2281 accgtgcatt tgacgtaaga aacttgcaat atcacgtttt aataagtttt gttcttcttc
     2341 agacattgta gagtaaattt caggtgttaa gaaagtacct ttaatttctt tgtaacctaa
     2401 aattgataat tcgtcagaaa tgtaagagta ttcaatgtta ggaattttaa cgtttgtttc
     2461 taagttagtg tttaagaagt tgtaaatagc tttttctttt gcgtaacctt tctttttgtt
     2521 agtagagaat ttagttttaa aaatgtattc gttgtttact aagtatgcaa cagaatcata
     2581 accagaacca ataatttcaa ttgagtcaac tttgaagtta tcaaagtagt gttcaattaa
     2641 gtatttcata gcttttacgt tagtagcgtt gtcatcataa cggtattcca ttaagtaaca
     2701 gtcttctttt ttaccttcgt gtaattcatg ttctggtaaa tcttcaataa tacggaaacc
     2761 acttttttga tatgcacgaa tagcacgagg gttgttttta tgaggatcta aaataactgc
     2821 gttagcgtta cgttcttttt taaggaattc gaaaattaat ttaatgtaac gtgtaccaat
     2881 acctttactc cagtagttag gttcaccaat gaattgatcc ataccgtaaa caatttcgtc
     2941 tgttttagga taatggtagt cagtgtataa ttcgtcatac attttgtaaa tttgaccata
     3001 accaattgga acgttgttat attcaataat tacacggaaa acttcgtctt cccatggttc
     3061 agtatagtgt tttttaagag attctaaagt gtattttttg tcacgaccac catagaattc
     3121 taaaacacgt tcatcagtta accattttaa cattaaaggg aagtcatcgt caattaatgt
     3181 acgaatacaa atttcgtttt caacaatgtt ttccattcga gaccactcat cgctctagaa
     3241 gcggccgcga attccagaaa tcatccttag cgaaagctaa ggattttttt tatctgaaat
     3301 tctgcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3361 cgtcaggtgg cactcgagtt gatcgggcac gtaagaggtt ccaactttca ccataatgaa
     3421 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     3481 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     3541 gaacat
//
