LOCUS       sequence-369409-        6767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty transposon system with constitutive bi-directional 
            promoter, expressing sfCherry11 fused to PDGFR through a (GGGGS)3 
            linker and the other side BFP and puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6767)
  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 6767)
  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-3xL-Delta-tag
FEATURES             Location/Qualifiers
     source          1..6767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     584..601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     975..994
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    993..1048
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1094..1113
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1213..1812)
                     /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(1296..1316)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1793..1812
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(1813..1869)
                     /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             complement(1879..2577)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        3209..4391
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     4339..4359
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4415..4424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     4421..4480
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4583..4612
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4616..4762
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(4811..4828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4817..5041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5473..5577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5578..5646,5647..6438)
                     /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     complement(5796..5815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6609..6767,1..430)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
       61 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      481 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      541 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      601 ggaagcggat ccctatacag ttgaagtcgg aagtttacat acaccttagc caaatacatt
      661 taaactcact ttttcacaat tcctgacatt taatcctagt aaaaattccc tgtcttaggt
      721 cagttaggat caccacttta ttttaagaat gtgaaatatc agaataatag tagagagaat
      781 gattcatttc agcttttatt tctttcatca cattcccagt gggtcagaag tttacataca
      841 ctcgactcct ctgcagaatg cggcgatgtt tcggtaaggg gtcccgagta gctagttcat
      901 ggcagccagc atatggcata tgttgccaaa ctctaaacca aatactcatt ctgatgtttt
      961 aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt ccaacacact
     1021 attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag gggcttcatg
     1081 atgtccccat aatttttggc agagggaaaa agatctcagt ggtatttgtg agccagggca
     1141 ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt gaattcacga
     1201 caggccttcg aatcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
     1261 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
     1321 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
     1381 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
     1441 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
     1501 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
     1561 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
     1621 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
     1681 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
     1741 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
     1801 gtactcggtc atggggccag gattctcctc gacgtcacca gcctgcttca gcaggctgaa
     1861 gttagtagct ccactgccgt tcagcttgtg ccccagtttg ctagggaggt cgcagtatct
     1921 ggccactgcc acctcgtgct gctcgacgta ggtctcgttg ttggcctcct tgattctttc
     1981 cagtctgtag tccacatagt agacgccagg catcttgagg ttcttagcgg gtttcttgga
     2041 tctatatgtg gtcttgatgt ttgcgatcag atggctcccg cccacgagct tcagggccat
     2101 gtcgtttctg ccttccaggc cgccgtcagc ggggtacagc gtctcggtga aggcctccca
     2161 gccgagtgtt ttcttctgca tcacagggcc gttggatgtg aagttcaccc ctctgatctt
     2221 gacgttgtag atgaggcagc cgtcctggag gctggtgtcc tgggtagcgg tcagcacgcc
     2281 cccgtcttcg tatgtggtga ctctctccca tgtgaagccc tcagggaagg actgcttgaa
     2341 gaagtcgggg atgccctggg tgtggttgat gaaggtcttg ctgccgtaga ggaagctagt
     2401 agccaggatg tcgaaggcga aggggagagg gccgccctcg accaccttga ttctcatggt
     2461 ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg cacttgaagt gatggttgtc
     2521 cacggtgccc tccatgtaca gcttcatgtg catgttctcc ttaatcagct cgctcatggt
     2581 ggcctcaggt gcagaggttt ctacagggaa agagagaaag gctacagtta gtttgctgtg
     2641 atgatgctaa gtgccgaggt cagatgtcta tggattgtac acggaatccc aacgcaggaa
     2701 ccaaaaaaca ctcacccaaa aaaatggcgc taaggccgag agaaaggcga agttccgtct
     2761 acggctatat accttgaaac aggggcggca gaagtgacgc aacgcggttg cgcggacggc
     2821 ttcctgtgtt ttcattggtc gaagcgttct gcgcgtgcgc agagattccc cagggaaggc
     2881 gggccctctt ccgaatgtcc aatccggaca ttcttccggt tggacctccg cggtcagcgc
     2941 tgtgcccggc gagagatcac gtggggcgcg gaggcggtgc tgctggggca cggccgtcca
     3001 gcctcggcgg ccatattttt gaggggctgt tcatctcgtt cacacgctct gtccgccatg
     3061 tttgtgagtg gaagcgccat taccccttca agcgactgaa ggctgcaggg cctctggtgg
     3121 cccgcatggg gagaccagac ccgccaggcc cgcctttccg cactcagtcc gggcttactt
     3181 tattttgtga gacagggtct cgcctagagg ctccggtgcc cgtcagtggg cagagcgcac
     3241 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     3301 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     3361 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     3421 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
     3481 gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc
     3541 ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag
     3601 ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa
     3661 tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt
     3721 tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag
     3781 atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc
     3841 agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt
     3901 agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc
     3961 cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc
     4021 ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg
     4081 agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca
     4141 cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt
     4201 ctttaggttg gggggagggg ttttatgcga tggagtttcc ccatactgag tgggtggaga
     4261 ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt
     4321 ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccattta
     4381 aggtgtcgtg aaaactaccc caagctggcc tctggccacc atggagacag acacactcct
     4441 gctatgggta ctgctgctct gggttccagg ttccactggt gacggctaca ccatcgtgga
     4501 gcagtacgag agagccgagg ccagacacag caccgcggga ggcggtggat cgggaggcgg
     4561 tggatcggga ggcggtggat cggaacaaaa actcatctca gaagaggatc tgaatgctgt
     4621 gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta aggtggtggt
     4681 gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca tcctcatcat
     4741 gctttggcag aagaagccac gttaggtcag gccaagcttg gggatcaatt ctctagagct
     4801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     5041 gcttctgagg cggaaagaac cagctgggcc tactagatag cggacccctt accgaaacat
     5101 cgccgcattc tgcagaggag tcgaacttag tgtatgtaaa cttctgaccc actggaattg
     5161 tgatacagtg aattataagt gaaataatct gtctgtaaac aattgttgga aaaatgactt
     5221 gtgtcatgca caaagtagat gtcctaactg acttgccaaa actattgttt gttaacaaga
     5281 aatttgtgga gtagttgaaa aacgagtttt aatgactcca acttaagtgt atgtaaactt
     5341 ccgacttcaa ctgtataggg atccaatttt aacaaaatat taacgcttac aatttcctga
     5401 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
     5461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5581 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5641 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5701 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5761 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5821 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5881 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5941 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6001 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6061 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6121 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6181 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6241 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6301 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6361 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6421 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgta
//
