LOCUS       sequence-322076-        2826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP for N-terminal fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2826)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 2826)
  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"
COMMENT     Sequence Label: pUPD2 GFP-NT (GB2236)
FEATURES             Location/Qualifiers
     source          1..2826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     508..527
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     755..773
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(777..820)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    823..844
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(862..864,865..867,868..1578)
                     /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(907..928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1168..1187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1515..1536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1622..1679
                     /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(1703..1720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1874..2462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1954..1973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2645..2739)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2760..2826,1..593))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
       61 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
      121 aaatcaaaac tggtgaaact cacccaggga ttggctgata cgaaaaacat attctcaata
      181 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
      241 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gtgatgaaaa cgtttcagtt
      301 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
      361 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
      421 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
      481 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
      541 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
      601 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
      661 ttatggtgaa agttggaacc tcttacgtgc ccgatcaact cgagtgccac ctgacgtcta
      721 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt
      781 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct
      841 agagcgatga gtggtctcgc catggtgagc aagggcgagg agctgttcac cggggtggtg
      901 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      961 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1021 ctgcccgtgc cctggcccac cctcgtgacc accttcagct acggcgtgca gtgcttcagc
     1081 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1141 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1201 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1261 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1321 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1381 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1441 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1501 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactcacggc
     1561 atggacgagc tgtacaagtc aatgtgagac cactcatcgc gactagtagc ggccgctgca
     1621 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     1681 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1861 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc
     2641 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     2701 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
     2761 tacgccccgc cctgccactc gtcgcagtac tgttgtaatt cattaagcat tctgccgaca
     2821 tggaag
//
