LOCUS       sequence-467921-        2996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes eGFP for ORBIT integration. High copy number..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2996)
  AUTHORS   Baughn A
  TITLE     Baughn lab ORBIT C-terminal eGFP tag plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2996)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pUMN105
FEATURES             Location/Qualifiers
     source          1..2996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(637..654)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(655..678)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(709..1419,1420..1422,1423..1425))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(751..772)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1100..1119
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1359..1380
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1428..1465
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      1518..1564
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgcgctagc
      601 ggccgcacta gtgcatgctc tagactcgag tcactagtgg tggtggtggt ggtgcttgtc
      661 gtcgtcgtcc ttgtagtcct ggaagtacag gttctcgccg ccgccgccct tgtacagctc
      721 gtccatgccc agggtgatgc ccgcggcggt cacgaactcc agcaggacca tgtggtcccg
      781 cttctcgttc gggtccttgc tcagcgccga ctgggtgctc aggtagtggt tgtccggcag
      841 caggacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcca gctggacgct
      901 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
      961 cgccatgatg tagacgttgt gcgagttgta gttgtactcc agcttgtggc ccaggatgtt
     1021 gccgtcttcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1081 gaacttgacc tcggcccggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1141 ctggacgtag ccctccggca tcgcgctctt gaagaagtcg tgctgcttca tgtggtcggg
     1201 gtaccgcgag aaacactgca cgccgtaggt cagggtggtc accagggtcg gccaggggac
     1261 cggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tcgcgtcgcc
     1321 ctcgccctcg cccgagacgc tgaacttgtg gccgttcacg tcgccgtcca gctcgaccag
     1381 gatgggcacg acgccggtga acagctcctc gcccttcgac accatcgatg atcctgacga
     1441 cggagaccgc cgtcgtcgac aagccaagct tatcgatgtc gacgtagtta actagcgtac
     1501 gatcgactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1561 ttatgccatc atggccgcgg tgatcagcta gagggggcgt caggcgccgg gggcggtgtc
     1621 cggcggcccc cagaggaact gcgccagttc ctccggatcg gtgaagccgg agagatccag
     1681 cggggtctcc tcgaacacct cgaagtcgtg caggaaggtg aaggcgagca gttcgcgggc
     1741 gaagtcctcg gtccgcttcc actgcgcccc gtcgagcagc gcggccagga tctcgcggtc
     1801 gccccggaag gcgttgagat gcagttgcac caggctgtag cgggagtctc ccgcatagac
     1861 gtcggtgaag tcgacgatcc cggtgacctc ggtcgcggcc aggtccacga agatgttggt
     1921 cccgtgcagg tcgccgtgga cgaaccgggg ttcgcggccg gccagcagcg tgtccacgtc
     1981 cggcagccag tcctccaggc ggtccagcag ccggggcgag aggtagcccc acccgcggtg
     2041 gtcctcgacg gtcgccgcgc ggcgttcccg cagcagttcc gggaagacct cggaatgggg
     2101 ggtgagcacg gtgttcccgg tcagcggcac cctgtgcagc cggccgagca cccggccgag
     2161 ttcgcgggcc agggcgagca gcgcgttccg gtcggtcgtg ccgtccatcg cggaccgcca
     2221 ggtggtgccg gtcatccggc tcatcaccag gtagggccac ggccaggctc cggtgccggg
     2281 ccgcagctcg ccgcggccga ggaggcgggg caccggcacc ggggcgtccg ccaggaccgc
     2341 gtacgcctcc gactccgacg cgaggctctc cggaccgcac cagtgctcgc cgaacagctt
     2401 gatcaccggg ccgggctcgc cgaccagtac ggggttggtg ctctcgccgg gcacccgcag
     2461 caccggcggc accggcagcc cgagctcctc cagggctcgg cgggccagcg gctcccagaa
     2521 ttcctggtcg ttccgcaggc tcgcgtagga atcatccgaa tcaatacggt cgagaagtaa
     2581 cagggattct tgtgtcacag cggacctcta ttcacagggt acgggccggc ttaattccgc
     2641 acggccggtc gcgacacggc ctgtccgcac cgcggatcag gcgttgacga tgacgggctg
     2701 gtcggccacg tcggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2821 gattgattta aaacttcagc attgccgttt ccgccagcca ctcatcgcag tactgttgta
     2881 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2941 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatgggg atcttc
//
