LOCUS       sequence-471486-        3135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a sfGFP cassette in the 3'UTR 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_5_001_3'UTR_Placeholder
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      420..514
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      697..1285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1186..1205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1439..1456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1480..1537)
                     /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."
     terminator      complement(1592..1619)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1635..1706)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1718..2431)
                     /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"
     RBS             2438..2449
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2624..2645
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      2648..2691
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2695..2713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2875..3135,1..399)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2941..2960)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc actctggagt
       61 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
      121 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtatcagcc
      181 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
      241 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
      301 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
      361 ttacaacagt actgcgacga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
      421 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
      481 cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctcgagctgt cagaccaagt
      541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
      601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
      661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
      721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1261 gtcagggggg cggagcctgt ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1381 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     1441 cgagtcagtg agcgaggaag cctgcataac gcgaagtaat cttttcggtt ttaaagaaaa
     1501 agggcagggt ggtgacacct tgcccttttt tgccggactg cagcggccgc tactagtcgc
     1561 gatgagtgtt ctcaagcgtg agacctataa acgcagaaag gcccacccga aggtgagcca
     1621 gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     1681 ttcgactgag cctttcgttt tatttgatgc ctggtcattt gtacagttca tccataccat
     1741 gcgtgatgcc cgctgcggtt acgaactcca gcagaaccat atgatcgcgt ttctcgttcg
     1801 gatctttaga cagaacgctt tgcgtgctca gatagtgatt gtctggcagc agaacaggac
     1861 catcaccgat tggagtgttt tgctggtagt gatcagccag ctgcacgctg ccatcctcca
     1921 cgttgtggcg aattttaaaa ttcgctttaa tgccattttt ttgtttatcg gcggtgatgt
     1981 aaacattgtg gctgttaaaa ttgtattcca gcttatggcc caggatattg ccgtcctctt
     2041 taaagtcaat gcctttcagc tcaatgcggt ttaccagggt atcgccttca aatttcactt
     2101 ccgcacgcgt tttgtacgtg ccgtcatcct taaaggaaat cgtgcgttcc tgcacatagc
     2161 cttccggcat ggcggacttg aagaagtcat gctgcttcat atggtccgga taacgagcaa
     2221 agcactgaac accataagtc agcgtcgtta ccagagtcgg ccaaggaacc ggcagtttac
     2281 cagtagtaca gatgaacttc agcgtcagtt taccattagt tgcgtcacct tcaccctcgc
     2341 cacgcacgga aaacttatga ccgttgacat caccatccag ttccaccaga atagggacga
     2401 caccagtgaa cagctcttcg cctttacgca tctagtattt ctcctctttc tctagtagct
     2461 aagatccgaa cctgccgttt cttgagttgc cgcgatgtta agaaaacatt cataaattaa
     2521 atgtgaattg ccgcacacat tattaaataa gatttacaaa atgttcaaaa tgacgcatga
     2581 aatcacgttt cactttcggt ctcaaagccg agactactca tcgctctaga agcggccgcg
     2641 aattccagaa atcatcctta gcgaaagcta aggatttttt ttatctgaaa ttctgcctcg
     2701 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2761 gcactcgagt tgatcgggca cgtaagaggt tccaactttc accataatga aataagatca
     2821 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
     2881 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     2941 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     3001 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     3061 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     3121 gtgatatggg atagt
//
