LOCUS       sequence-471476-        2190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLAG C-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2190)
  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 2190)
  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_5a_004_Flag-tag
FEATURES             Location/Qualifiers
     source          1..2190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(733..827)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(848..1507)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1422..1441
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1669..1687
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1691..1734)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1737..1758
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             1780..1803
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1807..1830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1834..1857
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1900..1957
                     /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(1981..1998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2152..2190,1..550))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
       61 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      361 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
      721 ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
      781 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
      841 tatcaaatta cgccccgccc tgccactcgt cgcagtactg ttgtaattca ttaagcattc
      901 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      961 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1021 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gatacgaaaa
     1081 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1141 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagtgatg
     1201 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     1261 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     1321 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     1381 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     1441 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     1501 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     1561 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aactcgagtg
     1621 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     1681 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat
     1741 tcgcggccgc ttctagagcg atgagtggtc tcggctttag attataaaga tgacgatgac
     1801 aaaggtgact ataaggatga tgatgataaa ggtgactaca aagacgacga tgataaaggg
     1861 tatgagacca ctcatcgcga ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt
     1921 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc
     1981 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2041 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag
     2161 gctccgcccc cctgacgagc atcacaaaaa
//
