LOCUS       sequence-433256-        2741 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid that encodes a sfGFP gene under a T7 promoter. Compatible 
            with PURExpress for in vitro transcription/translation. 
            Chloramphenicol resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2741)
  AUTHORS   Thompson, S, Zhang, Y, Yang, Z, Nichols,  LA, Fordyce, PM.
  TITLE     FACS-Sortable Microreactors for Ultra High-throughput Screening of 
            Catalysts in Biphasic Environments
  JOURNAL   bioRxiv.
REFERENCE   2  (bases 1 to 2741)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv."
COMMENT     Sequence Label: SMT000
FEATURES             Location/Qualifiers
     source          1..2741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(116..210)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(231..890)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGIMLNELQQYCDEWQGGA"
     primer_bind     805..824
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1028..1047
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1028..1046
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1071..1073,1074..1787)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1071..1784
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1101..1129)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1724..1744
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    1813..1833
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1834..1891
                     /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(1915..1932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2086..2674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2166..2185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
       61 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctcgagg cttggattct
      121 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttatg
      181 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
      241 ccctgccact catcgcagta ctgttgtaat tcattaagca ttatgccgac atggaagcca
      301 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      361 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
      421 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
      481 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
      541 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
      601 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
      661 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
      721 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
      781 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
      841 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
      901 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
      961 tggtgaaagt tggaacctct tacgtgcccg atcaactcga gtgccacctg acgtctaaga
     1021 aaccgaataa tacgactcac tatagggctt aagtataagg agatatacat atgagcaaag
     1081 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
     1141 ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga aaactcaccc
     1201 ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt gtcactactc
     1261 tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg catgactttt
     1321 tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg
     1381 ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg
     1441 agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa ctcgagtaca
     1501 actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta
     1561 acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac cattatcaac
     1621 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac
     1681 aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg
     1741 taactgctgc tgggattaca catggcatgg atgagctcta caaataacgc tgatagtgct
     1801 agtgtagatc gctactagta gcggccgctg cagtccggca aaaaagggca aggtgtcacc
     1861 accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct
     1921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg
     2101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2401 caacccggta agacacgact tatcgcaact ggcagcagcc actggtaaca ggattagcag
     2461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt t
//
