LOCUS       sequence-121277-        3440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobilizible plasmid with EGFP. OriT from the F plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  AUTHORS   Leonard J
  TITLE     Leonard Lab Conjugation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3440)
  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            "Unpublished"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: pShuttle
FEATURES             Location/Qualifiers
     source          1..3440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            91..380
                     /note="origin of transfer for the bacterial F plasmid 
                     (Frost et al., 1994)"
     primer_bind     504..522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(526..569)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    572..593
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    619..637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             656..667
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             674..1387
                     /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(704..732)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1327..1347
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1410..1481
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1497..1524
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1531..1551
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1552..1609
                     /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(1633..1650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1804..2392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1884..1903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2563..3354,3355..3423))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3186..3205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atttaagaac aggcagatgg ctaacatcca ttttttcatt tttccacctc tggtgacttt
       61 atccgtaaat aatttaaccc actccacaaa aaggctcaac aggttggtgg ttctcaccac
      121 caaaagcacc acaccccacg caaaaacaag tttttgctga ttttctttat aaatagagtg
      181 ttatgaaaaa ttagtttctc ttactctctt tatgatattt aaaaaagcgg tgtcggcgcg
      241 gctacaacaa cgcgccgaca ccgttttgta ggggtggtac tgactatttt tataaaaaac
      301 attattttat attaggggtg ctgctagcgg cgcggtgtgt ttttttatag gataccgcta
      361 ggggcgctgc tagcggtgcg tccctgtttg cattatgaat tttagtgttt cgaaattaac
      421 tttattttat gttcaaaaaa ggtaatctct aagtgccacc tgacgtctaa gaaaccatta
      481 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
      541 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagtccctat
      601 cagtgataga gattgacatc cctatcagtg atagagatac tgagcactac tagagaaaga
      661 ggagaaatac tagatgcgta aaggagaaga acttttcact ggagttgtcc caattcttgt
      721 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
      781 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
      841 atggccaaca cttgtcacta ctttcggtta tggtgttcaa tgctttgcga gatacccaga
      901 tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      961 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg
     1021 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
     1081 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
     1141 acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt
     1201 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     1261 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     1321 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     1381 atacaaataa taatactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1441 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1501 caccttcggg tgggcctttc tgcgtttata tactagtagc ggccgctgca gtccggcaaa
     1561 aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     1621 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1801 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2221 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3421 catactcttc ctttttcaat
//
