LOCUS       sequence-400023-        3923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORBIT integrating plasmid for C-terminal tagging with eGFP 
            containing a 3C protease cleavage site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3923)
  AUTHORS   Chen J, Fruhauf A, Fan C, Ponce J, Ueberheide B, Bhabha G, Ekiert D
  TITLE     Structure of an endogenous mycobacterial MCE lipid transporter.
  JOURNAL   Res Sq. 2023 Jan 10:rs.3.rs-2412186. doi: 
            10.21203/rs.3.rs-2412186/v1. Preprint.
  PUBMED    36711512
REFERENCE   2  (bases 1 to 3923)
  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: "Res Sq. 
            2023 Jan 10:rs.3.rs-2412186. doi: 10.21203/rs.3.rs-2412186/v1. 
            Preprint."
COMMENT     Sequence Label: pBEL2108
FEATURES             Location/Qualifiers
     source          1..3923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(629..648)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(2501..2547)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2600..2637
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             2649..2672
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2688..2690,2691..3401)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2730..2751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2991..3010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3338..3359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3432..3455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3456..3473
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(join(3699..3923,1..364))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3779..3798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
       61 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      181 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      361 tcaagaagat cctttgatct tttctacggg gtctgacgct catggaacta gagcatgaga
      421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatcggcacg taagaggttc
      481 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
      541 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
      601 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
      661 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
      721 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
      781 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
      841 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
      901 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
      961 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     1021 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1081 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1141 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1201 tgagtggctg gcggaaacgg caatgctgaa gttttaaatc aatctaaagt atatatgagt
     1261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1321 tatttcgttc atccatagtt gcctgactcc ccgacgtggc cgaccagccc gtcatcgtca
     1381 acgcctgatc cgcggtgcgg acaggccgtg tcgcgaccgg ccgtgcggaa ttaagccggc
     1441 ccgtaccctg tgaatagagg tccgctgtga cacaagaatc cctgttactt ctcgaccgta
     1501 ttgattcgga tgattcctac gcgagcctgc ggaacgacca ggaattctgg gagccgctgg
     1561 cccgccgagc cctggaggag ctcgggctgc cggtgccgcc ggtgctgcgg gtgcccggcg
     1621 agagcaccaa ccccgtactg gtcggcgagc ccggcccggt gatcaagctg ttcggcgagc
     1681 actggtgcgg tccggagagc ctcgcgtcgg agtcggaggc gtacgcggtc ctggcggacg
     1741 ccccggtgcc ggtgccccgc ctcctcggcc gcggcgagct gcggcccggc accggagcct
     1801 ggccgtggcc ctacctggtg atgagccgga tgaccggcac cacctggcgg tccgcgatgg
     1861 acggcacgac cgaccggaac gcgctgctcg ccctggcccg cgaactcggc cgggtgctcg
     1921 gccggctgca cagggtgccg ctgaccggga acaccgtgct caccccccat tccgaggtct
     1981 tcccggaact gctgcgggaa cgccgcgcgg cgaccgtcga ggaccaccgc gggtggggct
     2041 acctctcgcc ccggctgctg gaccgcctgg aggactggct gccggacgtg gacacgctgc
     2101 tggccggccg cgaaccccgg ttcgtccacg gcgacctgca cgggaccaac atcttcgtgg
     2161 acctggccgc gaccgaggtc accgggatcg tcgacttcac cgacgtctat gcgggagact
     2221 cccgctacag cctggtgcaa ctgcatctca acgccttccg gggcgaccgc gagatcctgg
     2281 ccgcgctgct cgacggggcg cagtggaagc ggaccgagga cttcgcccgc gaactgctcg
     2341 ccttcacctt cctgcacgac ttcgaggtgt tcgaggagac cccgctggat ctctccggct
     2401 tcaccgatcc ggaggaactg gcgcagttcc tctgggggcc gccggacacc gcccccggcg
     2461 cctgacgccc cctctagctg atcaccgcgg ccatgatggc ataaaacgaa aggcccagtc
     2521 tttcgactga gcctttcgtt ttatttgatg cctggcagtc gatcgtacgc tagttaacta
     2581 cgtcgacatc gataagcttg gcttgtcgac gacggcggtc tccgtcgtca ggatcatcgt
     2641 cgaattcttt ggaagttttg tttcaaggtc caactgctgc cgccgctgtg tcgaagggcg
     2701 aggagctgtt caccggcgtc gtgcccatcc tggtcgagct ggacggcgac gtgaacggcc
     2761 acaagttcag cgtctcgggc gagggcgagg gcgacgcgac ctacggcaag ctgaccctga
     2821 agttcatctg caccaccggc aagctgccgg tcccctggcc gaccctggtg accaccctga
     2881 cctacggcgt gcagtgtttc tcgcggtacc ccgaccacat gaagcagcac gacttcttca
     2941 agagcgcgat gccggagggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3001 actacaagac ccgggccgag gtcaagttcg agggcgacac cctggtgaac cgcatcgagc
     3061 tgaagggcat cgacttcaag gaagacggca acatcctggg ccacaagctg gagtacaact
     3121 acaactcgca caacgtctac atcatggcgg acaagcagaa gaacggcatc aaggtgaact
     3181 tcaagatccg ccacaacatc gaggacggca gcgtccagct ggccgaccac taccagcaga
     3241 acacccccat cggcgacggc cccgtcctgc tgccggacaa ccactacctg agcacccagt
     3301 cggcgctgag caaggacccg aacgagaagc gggaccacat ggtcctgctg gagttcgtga
     3361 ccgccgcggg catcaccctg ggcatggacg agctgtacaa gggcggcggc ggcgagaacc
     3421 tgtacttcca ggactacaag gacgacgacg acaagcacca ccaccaccac cactagtgac
     3481 tcgagtctag agcatgcact agtgcggccg ctagcggtac cagatctggg cccttaagtc
     3541 gcgttgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3601 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3661 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3721 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3781 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3841 cttaccggat acctgtccgc ctttctcccc tcgggaagcg tggcgctttc tcatagctca
     3901 cgctgtaggt atctcagttc ggt
//
