LOCUS       sequence-471521-        2120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' Connector 5 Short Forward in the M2 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_6_025_3C5SF
FEATURES             Location/Qualifiers
     source          1..2120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..26)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(698..792)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(813..1472)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1387..1406
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1634..1652
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1656..1699)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1702..1723
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1795..1852
                     /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(1876..1893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2047..2120,1..515))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
       61 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      121 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      361 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      661 tatatgagta aacttggtct gacagctcga ggcttggatt ctcaccaata aaaaacgccc
      721 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
      781 tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcgtcgcag
      841 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
      901 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
      961 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     1021 acccagggat tggctgatac gaaaaacata ttctcaataa accctttagg gaaataggcc
     1081 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     1141 tcgtggtatt cactccagag tgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     1201 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc
     1261 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     1321 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     1381 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     1441 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     1501 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     1561 cttacgtgcc cgatcaactc gagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1621 attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc
     1681 tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagcgatgag tggtctcgcg
     1741 ctggtaggag acgagactga gaccactcat cgcgactagt agcggccgct gcagtccggc
     1801 aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
     1861 cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2041 ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2101 gaggtggcga aacccgacag
//
