LOCUS       sequence-488472-        3135 bp    DNA     circular SYN 06-OCT-2025
DEFINITION  Encodes a sfGFP cassette in the promoter+5'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 Oct 6, 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_2-3_005_Promoter+5'UTR_Placeholder
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             160..171
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             178..891
                     /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      903..974
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      990..1017
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1072..1129
                     /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(1153..1170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1324..1912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1404..1423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2095..2189)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2210..2869)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2784..2803
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3031..3049
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3053..3096)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3099..3120
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagtgagac cgaaagtgaa acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt
       61 atttaataat gtgtgcggca attcacattt aatttatgaa tgttttctta acatcgcggc
      121 aactcaagaa acggcaggtt cggatcttag ctactagaga aagaggagaa atactagatg
      181 cgtaaaggcg aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
      241 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
      301 ctgacgctga agttcatctg tactactggt aaactgccgg ttccttggcc gactctggta
      361 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
      421 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
      481 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
      541 cgcattgagc tgaaaggcat tgactttaaa gaggacggca atatcctggg ccataagctg
      601 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
      661 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
      721 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
      781 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
      841 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgaccaggc
      901 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      961 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     1021 ataggtctca aatgtgagaa cactcatcgc gactagtagc ggccgctgca gtccggcaaa
     1081 aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     1141 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1321 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1741 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2041 taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc ttggattctc
     2101 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     2161 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     2221 cctgccactc gtcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     2281 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     2341 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     2401 caaaactggt gaaactcacc cagggattgg ctgatacgaa aaacatattc tcaataaacc
     2461 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     2521 gaaactgccg gaaatcgtcg tggtattcac tccagagtga tgaaaacgtt tcagtttgct
     2581 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     2641 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     2701 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     2761 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     2821 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     2881 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2941 ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag tgccacctga cgtctaagaa
     3001 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     3061 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag
     3121 cgatgagtag tctcg
//
