LOCUS       sequence-471433-        3135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a sfGFP cassette in the promoter 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_2_001_Promoter_placeholder
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      582..676
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      859..1447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1348..1367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1601..1618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1642..1699)
                     /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(1754..1781)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1797..1868)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1880..2593)
                     /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             2600..2611
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2786..2807
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      2810..2853
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2857..2875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(3037..3135,1..561)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3103..3122)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
       61 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
      121 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
      181 gttttccatg agcaaactga aacgttttca tcactctgga gtgaatacca cgacgatttc
      241 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      301 ttccctaaag ggtttattga gaatatgttt ttcgtatcag ccaatccctg ggtgagtttc
      361 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      421 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      481 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgac
      541 gagtggcagg gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc
      601 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
      661 ttttattggt gagaatccaa gcctcgagct gtcagaccaa gtttactcat atatacttta
      721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      961 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1441 gtggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1501 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1561 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1621 agcctgcata acgcgaagta atcttttcgg ttttaaagaa aaagggcagg gtggtgacac
     1681 cttgcccttt tttgccggac tgcagcggcc gctactagtc gcgatgagtg ttctcaagta
     1741 tgagacctat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgact ctagtagaga
     1801 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     1861 tttatttgat gcctggtcat ttgtacagtt catccatacc atgcgtgatg cccgctgcgg
     1921 ttacgaactc cagcagaacc atatgatcgc gtttctcgtt cggatcttta gacagaacgc
     1981 tttgcgtgct cagatagtga ttgtctggca gcagaacagg accatcaccg attggagtgt
     2041 tttgctggta gtgatcagcc agctgcacgc tgccatcctc cacgttgtgg cgaattttaa
     2101 aattcgcttt aatgccattt ttttgtttat cggcggtgat gtaaacattg tggctgttaa
     2161 aattgtattc cagcttatgg cccaggatat tgccgtcctc tttaaagtca atgcctttca
     2221 gctcaatgcg gtttaccagg gtatcgcctt caaatttcac ttccgcacgc gttttgtacg
     2281 tgccgtcatc cttaaaggaa atcgtgcgtt cctgcacata gccttccggc atggcggact
     2341 tgaagaagtc atgctgcttc atatggtccg gataacgagc aaagcactga acaccataag
     2401 tcagcgtcgt taccagagtc ggccaaggaa ccggcagttt accagtagta cagatgaact
     2461 tcagcgtcag tttaccatta gttgcgtcac cttcaccctc gccacgcacg gaaaacttat
     2521 gaccgttgac atcaccatcc agttccacca gaatagggac gacaccagtg aacagctctt
     2581 cgcctttacg catctagtat ttctcctctt tctctagtag ctaagatccg aacctgccgt
     2641 ttcttgagtt gccgcgatgt taagaaaaca ttcataaatt aaatgtgaat tgccgcacac
     2701 attattaaat aagatttaca aaatgttcaa aatgacgcat gaaatcacgt ttcactttcg
     2761 gtctcactcc cgagactact catcgctcta gaagcggccg cgaattccag aaatcatcct
     2821 tagcgaaagc taaggatttt ttttatctga aattctgcct cgtgatacgc ctatttttat
     2881 aggttaatgt catgataata atggtttctt agacgtcagg tggcactcga gttgatcggg
     2941 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     3001 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     3061 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     3121 gctcaatgta cctat
//
