LOCUS       sequence-447499-        2808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 Entry vector"" ORF - eGFP                         "".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2808)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 2808)
  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: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pBP-ORF-eGFP
FEATURES             Location/Qualifiers
     source          1..2808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..717
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(34..62)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     657..677
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      754..811
                     /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(835..852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1006..1594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1086..1105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1777..1871)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1892..2551)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2466..2485
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2713..2731
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2735..2778)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2781..2802
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 catatgcgta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
       61 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
      121 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
      181 cttgtcacta ctttcggtta tggtgttcaa tgctttgcga gatacccaga tcatatgaaa
      241 cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
      301 ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt
      361 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
      421 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
      481 ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
      541 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
      601 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga tcacatggtc
      661 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa
      721 tcgaagagac ccatgccgtc tcattagctg cagtccggca aaaaagggca aggtgtcacc
      781 accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct
      841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg
     1021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1741 atatgagtaa acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg
     1801 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     1861 caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt
     1921 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     1981 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     2041 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     2101 cccagggatt ggctgacacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     2161 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     2221 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     2281 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     2341 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     2401 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     2461 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     2521 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     2581 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     2641 ttacgtgccc gatcaactcg agtgccacct gacgtctaag aaaccattat tatcatgaca
     2701 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     2761 ttcgctaagg atgatttctg gaattcgcgg ccgcttctag aggtctca
//
