LOCUS       sequence-471520-        2120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' Connector 4 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_024_3C4SF
FEATURES             Location/Qualifiers
     source          1..2120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     229..246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(270..327)
                     /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    399..420
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      423..466
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(470..488)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             650..1309
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(716..735)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1330..1424
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(1607..2120,1..75)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2096..2115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
       61 cggagcctgt ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      121 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      181 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      241 agcgaggaag cctgcataac gcgaagtaat cttttcggtt ttaaagaaaa agggcagggt
      301 ggtgacacct tgcccttttt tgccggactg cagcggccgc tactagtcgc gatgagtggt
      361 ctcagtctcg tctcccttga gcgcgagacc actcatcgct ctagaagcgg ccgcgaattc
      421 cagaaatcat ccttagcgaa agctaaggat tttttttatc tgaaattctg cctcgtgata
      481 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      541 cgagttgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
      601 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
      661 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      721 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
      781 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
      841 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
      901 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
      961 actctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1021 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1081 cgtatcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1141 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1201 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1261 gcttaatgaa ttacaacagt actgcgacga gtggcagggc ggggcgtaat ttgatatcga
     1321 gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1381 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctcgagctgt
     1441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2101 acgcctggta tctttatagt
//
