LOCUS       sequence-471421-        2120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' Connector 4 Short Forward 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_022_5C4SF
FEATURES             Location/Qualifiers
     source          1..2120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     810..828
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(832..875)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    878..899
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      971..1028
                     /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(1052..1069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1223..1811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1303..1322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1994..2088)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2109..2120,1..648))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ctgccactcg tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
       61 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
      121 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
      181 aaaactggtg aaactcaccc agggattggc tgatacgaaa aacatattct caataaaccc
      241 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
      301 aaactgccgg aaatcgtcgt ggtattcact ccagagtgat gaaaacgttt cagtttgctc
      361 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
      421 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
      481 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
      541 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
      601 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
      661 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
      721 gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa
      781 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
      841 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagagc
      901 gatgagtggt ctcgaacacg tctcgcaagg gagtgagacc actcatcgcg actagtagcg
      961 gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac
     1021 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
     1081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1201 aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa
     1261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1981 gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     2041 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     2101 atatcaaatt acgccccgcc
//
