LOCUS       sequence-372902-        6788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty transposon system with constitutive bi-directional 
            promoter, one side expressing SpyTag003-sfCherry11 fused to PDGFR 
            and the other side expressing BFP and puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6788)
  AUTHORS   Moghimianavval H, Patel C, Mohapatra S, Hwang SW, Kayikcioglu T, 
            Bashirzadeh Y, Liu AP, Ha T
  TITLE     Engineering Functional Membrane-Membrane Interfaces by InterSpy.
  JOURNAL   Small. 2022 May 26:e2202104. doi: 10.1002/smll.202202104.
  PUBMED    35618485
REFERENCE   2  (bases 1 to 6788)
  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: "Small. 2022
            May 26:e2202104. doi: 10.1002/smll.202202104."
COMMENT     Sequence Label: InterTag
FEATURES             Location/Qualifiers
     source          1..6788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             565..1263
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1273..1329
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1330..1929
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1330..1349)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1826..1846
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2029..2048)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2094..2149
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2148..2167)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2541..2558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2712..3300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2792..2811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3471..4262,4263..4331))
                     /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     4094..4113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4332..4436)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4868..5092
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5081..5098
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5147..5293)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(5297..5326)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(5396..5443)
                     /codon_start=1
                     /product="improved SpyTag variant that reacts with 
                     SpyCatcher003 at a rate approaching the diffusion limit 
                     (Keeble et al., 2019)"
                     /label=SpyTag003
                     /translation="RGVPHIVMVDAYKRYK"
     sig_peptide     complement(5450..5509)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      5506..5515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5539..6721)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(5571..5591)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ggtctggtct ccccatgcgg gccaccagag gccctgcagc cttcagtcgc ttgaaggggt
       61 aatggcgctt ccactcacaa acatggcgga cagagcgtgt gaacgagatg aacagcccct
      121 caaaaatatg gccgccgagg ctggacggcc gtgccccagc agcaccgcct ccgcgcccca
      181 cgtgatctct cgccgggcac agcgctgacc gcggaggtcc aaccggaaga atgtccggat
      241 tggacattcg gaagagggcc cgccttccct ggggaatctc tgcgcacgcg cagaacgctt
      301 cgaccaatga aaacacagga agccgtccgc gcaaccgcgt tgcgtcactt ctgccgcccc
      361 tgtttcaagg tatatagccg tagacggaac ttcgcctttc tctcggcctt agcgccattt
      421 ttttgggtga gtgttttttg gttcctgcgt tgggattccg tgtacaatcc atagacatct
      481 gacctcggca cttagcatca tcacagcaaa ctaactgtag cctttctctc tttccctgta
      541 gaaacctctg cacctgaggc caccatgagc gagctgatta aggagaacat gcacatgaag
      601 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
      661 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      721 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
      781 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
      841 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      901 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
      961 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
     1021 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
     1081 aacatcaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
     1141 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
     1201 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctg
     1261 aacggcagtg gagctactaa cttcagcctg ctgaagcagg ctggtgacgt cgaggagaat
     1321 cctggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1381 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1441 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1501 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     1561 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     1621 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     1681 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     1741 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     1801 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     1861 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     1921 ggtgcctgat tcgaaggcct gtcgtgaatt cactcctcag gtgcaggctg cctatcagaa
     1981 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     2041 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     2101 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     2161 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     2221 atatgccata tgctggctgc catgaactag ctactcggga ccccttaccg aaacatcgcc
     2281 gcattctgca gaggagtcga gtgtatgtaa acttctgacc cactgggaat gtgatgaaag
     2341 aaataaaagc tgaaatgaat cattctctct actattattc tgatatttca cattcttaaa
     2401 ataaagtggt gatcctaact gacctaagac agggaatttt tactaggatt aaatgtcagg
     2461 aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg tatgtaaact tccgacttca
     2521 actgtatagg gatccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4441 tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4501 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattggatcc ctatacagtt
     4561 gaagtcggaa gtttacatac acttaagttg gagtcattaa aactcgtttt tcaactactc
     4621 cacaaatttc ttgttaacaa acaatagttt tggcaagtca gttaggacat ctactttgtg
     4681 catgacacaa gtcatttttc caacaattgt ttacagacag attatttcac ttataattca
     4741 ctgtatcaca attccagtgg gtcagaagtt tacatacact aagttcgact cctctgcaga
     4801 atgcggcgat gtttcggtaa ggggtccgct atctagtagg cccagctggt tctttccgcc
     4861 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     4921 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     4981 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     5041 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     5101 gatcagcgag ctctagagaa ttgatcccca agcttggcct gacctaacgt ggcttcttct
     5161 gccaaagcat gatgaggatg ataagggaga tgatggtgag caccaccagg gccaggatgg
     5221 ctgagatcac caccacctta aagggcaagg agtgtggcac cacgatgacc tcctgcgtgt
     5281 cctggcccac agcattcaga tcctcttctg agatgagttt ttgttcggcg gtgctgtgtc
     5341 tggcctcggc tctctcgtac tgctccacga tggtgtagcc tgaaccacct cccgatttat
     5401 agcgtttgta ggcgtccacc ataacaatat gaggaacgcc acgtccgtca ccagtggaac
     5461 ctggaaccca gagcagcagt acccatagca ggagtgtgtc tgtctccatg gtggccagag
     5521 gccagcttgg ggtagttttc acgacacctt aaatggaaga aaaaaacttt gaaccactgt
     5581 ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta
     5641 catcaagtgc caagctggcc taacttcagt ctccacccac tcagtatggg gaaactccat
     5701 cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg agaactaatc
     5761 gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg
     5821 aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc gccgcgtcct
     5881 ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc acgcaactgg
     5941 tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg cgaggccagg
     6001 caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg gccgcgctcg
     6061 caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc
     6121 cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac tatcttgcca
     6181 gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc
     6241 agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaagcca
     6301 ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca
     6361 cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg ggcgtggaag
     6421 taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca
     6481 cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     6541 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     6601 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
     6661 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     6721 ctctaggcga gaccctgtct cacaaaataa agtaagcccg gactgagtgc ggaaaggcgg
     6781 gcctggcg
//
