LOCUS       sequence-471425-        2120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' Connector 2 Short Reverse in the M1 position used to dictate 
            position for lvl2 assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2120)
  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 2120)
  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_G_0_1_009_5C2SR
FEATURES             Location/Qualifiers
     source          1..2120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      183..771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     672..691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     925..942
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(966..1023)
                     /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    1095..1116
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1119..1162
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1166..1184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1346..2005
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1412..1431)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2026..2120
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ccaagcctcg agctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
       61 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      121 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      181 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      241 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      301 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      361 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      421 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      481 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      541 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      601 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      661 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      721 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctgtggaa aaacgccagc
      781 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      841 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      901 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcctg cataacgcga
      961 agtaatcttt tcggttttaa agaaaaaggg cagggtggtg acaccttgcc cttttttgcc
     1021 ggactgcagc ggccgctact agtcgcgatg agtggtctca ctccaatgcg agacgtgttc
     1081 gagaccactc atcgctctag aagcggccgc gaattccaga aatcatcctt agcgaaagct
     1141 aaggattttt tttatctgaa attctgcctc gtgatacgcc tatttttata ggttaatgtc
     1201 atgataataa tggtttctta gacgtcaggt ggcactcgag ttgatcgggc acgtaagagg
     1261 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     1321 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     1381 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1441 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1501 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1561 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1621 caccgttttc catgagcaaa ctgaaacgtt ttcatcactc tggagtgaat accacgacga
     1681 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1741 ctatttccct aaagggttta ttgagaatat gtttttcgta tcagccaatc cctgggtgag
     1801 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1861 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1921 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1981 cgacgagtgg cagggcgggg cgtaatttga tatcgagctc gcttggactc ctgttgatag
     2041 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     2101 cgttttttat tggtgagaat
//
