LOCUS       sequence-471478-        3135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a sfGFP cassette in the CDS position used for placeholder 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3135)
  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 3135)
  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_4_001_CDS_Placeholder
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(535..629)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(650..1309)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1224..1243
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1471..1489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1493..1536)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1539..1560
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             1735..1746
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1753..2466
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2478..2549
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2565..2592
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2647..2704
                     /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(2728..2745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2899..3135,1..352))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2979..2998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
       61 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc ttggattctc
      541 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
      601 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
      661 cctgccactc gtcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
      721 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
      781 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
      841 caaaactggt gaaactcacc cagggattgg ctgatacgaa aaacatattc tcaataaacc
      901 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
      961 gaaactgccg gaaatcgtcg tggtattcac tccagagtga tgaaaacgtt tcagtttgct
     1021 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     1081 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     1141 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     1201 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     1261 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     1321 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     1381 ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag tgccacctga cgtctaagaa
     1441 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     1501 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag
     1561 cgatgagtag tctcgaatgt gagaccgaaa gtgaaacgtg atttcatgcg tcattttgaa
     1621 cattttgtaa atcttattta ataatgtgtg cggcaattca catttaattt atgaatgttt
     1681 tcttaacatc gcggcaactc aagaaacggc aggttcggat cttagctact agagaaagag
     1741 gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
     1801 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
     1861 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggttcct
     1921 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
     1981 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
     2041 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
     2101 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagagga cggcaatatc
     2161 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
     2221 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
     2281 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
     2341 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     2401 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     2461 tacaaatgac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2521 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2581 ggcctttctg cgtttatagg tctcagcttt gagaacactc atcgcgacta gtagcggccg
     2641 ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     2701 aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcg
//
