LOCUS       sequence-422615-        4160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mariner transposon expressing sfGFP with kan selection casette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4160)
  AUTHORS   English MA, Alcantar MA, Collins JJ
  TITLE     A self-propagating, barcoded transposon system for the dynamic 
            rewiring of genomic networks.
  JOURNAL   Mol Syst Biol. 2023 Jun 12;19(6):e11398. doi: 
            10.15252/msb.202211398. Epub 2023 Mar 27.
  PUBMED    36970845
REFERENCE   2  (bases 1 to 4160)
  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: "Mol Syst 
            Biol."; date: "2023-06-12"; volume: "19(6):e11398. doi"; pages: " 
            10.15252/msb.202211398. Epub 2023 Mar 27"
COMMENT     Sequence Label: pDonor_Tn[sfGFP]
FEATURES             Location/Qualifiers
     source          1..4160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(18..809,810..878))
                     /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     641..660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(879..983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1051..1069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1184..1218
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_RNA        1229..1278
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(1321..1323,1324..2037)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1321..2034
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1351..1379)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1974..1994
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2348..2367)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2359..3174
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2359..3174
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2432..2451)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     oriT            3905..4014
                     /note="incP origin of transfer"
ORIGIN
        1 ccctcgaatt gacgcgttta ccaatgctta atcagtgagg cacctatctc agcgatctgt
       61 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
      601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
      661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
      721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
      781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
      841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
      901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
      961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1021 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcac
     1081 gcgtcctcgg tacctaacag gttggatgat aagtccccgg tcttcgtatg ccgtcttctg
     1141 cttggcgcgc cctcgacgtc tccagcttcg tacgcggccg gcctttacag ctagctcagt
     1201 cctaggtatt atgctagcct cgagttagct gtcaccggat gtgctttccg gtctgatgag
     1261 tccgtgagga cgaaacagcc tctacaaata attttgttta atcacacagg aaagtactag
     1321 atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     1381 gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga
     1441 aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt
     1501 gtcactactc tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg
     1561 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
     1621 aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1681 aatcgtatcg agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa
     1741 cttgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     1801 atcaaagcta acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac
     1861 cattatcaac aaaatactcc aatcggcgat ggccctgtcc ttttaccaga caaccattac
     1921 ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     1981 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaataagga
     2041 tccttcagcc aaaaaactta agaccgccgg tcttgtccac taccttgcag taatgcggtg
     2101 gacaggatcg gcggttttct tttctcttct caactcggta ccaaagacga acaataagac
     2161 gctgaaaagc gtcttttttc gttttggtcc atgcctttaa ttaatctaga ctgatccttc
     2221 aactcagcaa aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt
     2281 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     2341 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc caggccgcga
     2401 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     2461 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     2521 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     2581 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     2641 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
     2701 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     2761 cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca
     2821 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     2881 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     2941 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     3001 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     3061 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     3121 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     3181 ttggttaatt ggttgtaaca ccaattggga aacacagaaa aaagcccgca cctgacagtg
     3241 cgggcttttt ttttcgacca aaggctcggt accaaattcc agaaaagaca cccgaaaggg
     3301 tgttttttcg ttttggtccc ccgggaacct gcagggcgcg ccaagcagaa gacggcatac
     3361 gaagaccggg gacttatcat ccaacctgtt aggatcagtg agcgcaacgg aatctaattc
     3421 ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc
     3481 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa
     3541 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg
     3601 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct
     3661 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg
     3721 agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt gaactgctga
     3781 tcttcagatc ctctacgccg gacgcatcgt ggccggatca attccgtgat aggtgggctg
     3841 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     3901 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     3961 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     4021 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     4081 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     4141 gcggaaaacg gaattgatcc
//
