LOCUS       sequence-314897-        2778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent protein coding sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2778)
  AUTHORS   Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , 
            Nakayama, Naomi
  TITLE     Mobius Assembly for Plant Systems highlights promoter-terminator 
            interaction in gene regulation
  JOURNAL   bioRxiv, March 31, 2021
REFERENCE   2  (bases 1 to 2778)
  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, 
            March 31, 2021"
COMMENT     Sequence Label: lvl0 eGFP CDS
FEATURES             Location/Qualifiers
     source          1..2778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..77
                     /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(101..118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(272..860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(352..371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1043..1137)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1158..1817)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1732..1751
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1979..1997
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2001..2044)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(2061..2063,2064..2066,2067..2778,1..2)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2106..2127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2367..2386)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2714..2735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aagcttcgag accctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt
       61 ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc
      121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc cctgacgagc
      301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1021 ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     1081 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1141 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     1201 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     1261 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     1321 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     1381 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     1441 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     1501 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     1561 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     1621 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     1681 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     1741 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     1801 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     1861 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     1921 aactcgagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     1981 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     2041 tttctggaat tcggtctcaa atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2101 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2161 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2221 tgcccgtgcc ctggcccacc ctcgtgacca ccttcagcta cggcgtgcag tgcttcagcc
     2281 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2341 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2401 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2461 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2521 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2581 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2641 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2701 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actcacggca
     2761 tggacgagct gtacaagt
//
