LOCUS       sequence-471472-        2190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLAG N-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_3b_026_Flag-tag
FEATURES             Location/Qualifiers
     source          1..2190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..43
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(67..124)
                     /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."
     CDS             complement(168..191)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(195..218)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(222..245)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    266..287
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      290..333
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(337..355)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             517..1176
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(583..602)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1197..1291
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1474..2062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1963..1982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcct gcataacgcg
       61 aagtaatctt ttcggtttta aagaaaaagg gcagggtggt gacaccttgc ccttttttgc
      121 cggactgcag cggccgctac tagtcgcgat gagtggtctc acattccttt atcatcgtcg
      181 tctttgtagt cacctttatc atcatcatcc ttatagtcac ctttgtcatc gtcatcttta
      241 taatccatgg cgagaccact catcgctcta gaagcggccg cgaattccag aaatcatcct
      301 tagcgaaagc taaggatttt ttttatctga aattctgcct cgtgatacgc ctatttttat
      361 aggttaatgt catgataata atggtttctt agacgtcagg tggcactcga gttgatcggg
      421 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
      481 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
      541 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
      601 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
      661 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
      721 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
      781 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcact ctggagtgaa
      841 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
      901 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt atcagccaat
      961 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1021 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1081 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     1141 caacagtact gcgacgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact
     1201 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     1261 ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc gagctgtcag accaagttta
     1321 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2041 aggggggcgg agcctgtgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2161 tattaccgcc tttgagtgag ctgataccgc
//
