LOCUS       sequence-369616-        7742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human 41BBL and CD86 in a Sleeping Beauty transposon 
            vector for use in artificial antigen presenting cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7742)
  AUTHORS   Dobson LJ, Saunderson SC, Smith-Bell SW, McLellan AD
  TITLE     Sleeping Beauty kit sets provide rapid and accessible generation of 
            artificial antigen-presenting cells for natural killer cell 
            expansion.
  JOURNAL   Immunol Cell Biol. 2023 Aug 16. doi: 10.1111/imcb.12679.
  PUBMED    37585342
REFERENCE   2  (bases 1 to 7742)
  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: "Immunol 
            Cell Biol. 2023 Aug 16. doi: 10.1111/imcb.12679."
COMMENT     Sequence Label: pSBbi 41BBL-CD86
FEATURES             Location/Qualifiers
     source          1..7742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..73)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(973..1184)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1833..1842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2610..2666
                     /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             3663..3719
                     /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"
     primer_bind     complement(4419..4438)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4484..4539
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4538..4557)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4931..4948)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5102..5690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5182..5201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5861..6652,6653..6721))
                     /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     6484..6503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6722..6826)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7258..7482
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7471..7488
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      7728..7737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aggccagctt ggggtagttt tcacgacacc ttaaatggaa gaaaaaaact ttgaaccact
       61 gtctgaggct tgagaatgaa ccaagatcca aactcaaaaa gggcaaattc caaggagaat
      121 tacatcaagt gccaagctgg cctaacttca gtctccaccc actcagtatg gggaaactcc
      181 atcgcataaa acccctcccc ccaacctaaa gacgacgtac tccaaaagct cgagaactaa
      241 tcgaggtgcc tggacggcgc ccggtactcc gtggagtcac atgaagcgac ggctgaggac
      301 ggaaaggccc ttttcctttg tgtgggtgac tcacccgccc gctctcccga gcgccgcgtc
      361 ctccattttg agctccctgc agcagggccg ggaagcggcc atctttccgc tcacgcaact
      421 ggtgccgacc gggccagcct tgccgcccag ggcggggcga tacacggcgg cgcgaggcca
      481 ggcaccagag caggccggcc agcttgagac tacccccgtc cgattctcgg tggccgcgct
      541 cgcaggcccc gcctcgccga acatgtgcgc tgggacgcac gggccccgtc gccgcccgcg
      601 gccccaaaaa ccgaaatacc agtgtgcaga tcttggcccg catttacaag actatcttgc
      661 cagaaaaaaa gcgtcgcagc aggtcatcaa aaattttaaa tggctagaga cttatcgaaa
      721 gcagcgagac aggcgcgaag gtgccaccag attcgcacgc ggcggcccca gcgcccaagc
      781 caggcctcaa ctcaagcacg aggcgaaggg gctccttaag cgcaaggcct cgaactctcc
      841 cacccacttc caacccgaag ctcgggatca agaatcacgt actgcagcca ggggcgtgga
      901 agtaattcaa ggcacgcaag ggccataacc cgtaaagagg ccaggcccgc gggaaccaca
      961 cacggcactt acctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
     1021 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
     1081 atcactttcc cagtttaccc cgcgccacct tctctaggca ccggttcaat tgccgacccc
     1141 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga
     1201 gcctctaggc gagaccctgt ctcacaaaat aaagtaagcc cggactgagt gcggaaaggc
     1261 gggcctggcg ggtctggtct ccccatgcgg gccaccagag gccctgcagc cttcagtcgc
     1321 ttgaaggggt aatggcgctt ccactcacaa acatggcgga cagagcgtgt gaacgagatg
     1381 aacagcccct caaaaatatg gccgccgagg ctggacggcc gtgccccagc agcaccgcct
     1441 ccgcgcccca cgtgatctct cgccgggcac agcgctgacc gcggaggtcc aaccggaaga
     1501 atgtccggat tggacattcg gaagagggcc cgccttccct ggggaatctc tgcgcacgcg
     1561 cagaacgctt cgaccaatga aaacacagga agccgtccgc gcaaccgcgt tgcgtcactt
     1621 ctgccgcccc tgtttcaagg tatatagccg tagacggaac ttcgcctttc tctcggcctt
     1681 agcgccattt ttttgggtga gtgttttttg gttcctgcgt tgggattccg tgtacaatcc
     1741 atagacatct gacctcggca cttagcatca tcacagcaaa ctaactgtag cctttctctc
     1801 tttccctgta gaaacctctg cacctgaggg cggccaccat ggaatacgcc tctgacgctt
     1861 cactggaccc cgaagccccg tggcctcccg cgccccgcgc tcgcgcctgc cgcgtactgc
     1921 cttgggccct ggtcgcgggg ctgctgctgc tgctgctgct cgctgccgcc tgcgccgtct
     1981 tcctcgcctg cccctgggcc gtgtccgggg ctcgcgcctc gcccggctcc gcggccagcc
     2041 cgagactccg cgagggtccc gagctttcgc ccgacgatcc cgccggcctc ttggacctgc
     2101 ggcagggcat gtttgcgcag ctggtggccc aaaatgttct gctgatcgat gggcccctga
     2161 gctggtacag tgacccaggc ctggcaggcg tgtccctgac ggggggcctg agctacaaag
     2221 aggacacgaa ggagctggtg gtggccaagg ctggagtcta ctatgtcttc tttcaactag
     2281 agctgcggcg cgtggtggcc ggcgagggct caggctccgt ttcacttgcg ctgcacctgc
     2341 agccactgcg ctctgctgct ggggccgccg ccctggcttt gaccgtggac ctgccacccg
     2401 cctcctccga ggctcggaac tcggccttcg gtttccaggg ccgcttgctg cacctgagtg
     2461 ccggccagcg cctgggcgtc catcttcaca ctgaggccag ggcacgccat gcctggcagc
     2521 ttacccaggg cgccacagtc ttgggactct tccgggtgac ccccgaaatc ccagccggac
     2581 tcccttcacc gaggtcggaa ggatccggag ctactaactt cagcctgctg aagcaggctg
     2641 gtgacgtcga ggagaatcct ggccccatgg atccccagtg cactatggga ctgagtaaca
     2701 ttctctttgt gatggccttc ctgctctctg gtgctgctcc tctgaagatt caagcttatt
     2761 tcaatgagac tgcagacctg ccatgccaat ttgcaaactc tcaaaaccaa agcctgagtg
     2821 agctagtagt attttggcag gaccaggaaa acttggttct gaatgaggta tacttaggca
     2881 aagagaaatt tgacagtgtt cattccaagt atatgggccg cacaagtttt gattcggaca
     2941 gttggaccct gagacttcac aatcttcaga tcaaggacaa gggcttgtat caatgtatca
     3001 tccatcacaa aaagcccaca ggaatgattc gcatccacca gatgaattct gaactgtcag
     3061 tgcttgctaa cttcagtcaa cctgaaatag taccaatttc taatataaca gaaaatgtgt
     3121 acataaattt gacctgctca tctatacacg gttacccaga acctaagaag atgagtgttt
     3181 tgctaagaac caagaattca actatcgagt atgatggtat tatgcagaaa tctcaagata
     3241 atgtcacaga actgtacgac gtttccatca gcttgtctgt ttcattccct gatgttacga
     3301 gcaatatgac catcttctgt attctggaaa ctgacaagac gcggctttta tcttcacctt
     3361 tctctataga gcttgaggac cctcagcctc ccccagacca cattccttgg attacagctg
     3421 tacttccaac agttattata tgtgtgatgg ttttctgtct aattctatgg aaatggaaga
     3481 agaagaagcg gcctcgcaac tcttataaat gtggaaccaa cacaatggag agggaagaga
     3541 gtgaacagac caagaaaaga gaaaaaatcc atatacctga aagatctgat gaagcccagc
     3601 gtgtttttaa aagttcgaag acatcttcat gcgacaaaag tgatacatgt tttggatccg
     3661 gagctactaa cttcagcctg ctgaagcagg ctggtgacgt cgaggagaat cctggcccca
     3721 tgacagaata caagcctaca gttagactgg caacacgcga tgatgttccc agggcggttc
     3781 ggacacttgc tgctgctttc gccgattatc cggccacgcg acacactgtg gacccagata
     3841 ggcacatcga gagagtgacg gagctgcaag agctgttttt gactagggtt ggacttgata
     3901 tcggaaaagt ttgggttgct gatgatggtg ctgcggtagc ggtctggaca actcctgagt
     3961 cagtggaggc cggagcggtc ttcgctgaga tcggtcctcg catggccgaa ttgagcggaa
     4021 gtcgattggc cgctcagcag caaatggagg gtcttctggc accccacagg cctaaggagc
     4081 ctgcgtggtt tcttgcaaca gtgggcgtgt ctccagatca tcaggggaaa ggtcttggtt
     4141 ctgcagtagt gctgcccggt gtagaggcag cggagagggc aggcgtacca gcgttcttgg
     4201 aaacctcagc cccgagaaac ctgcctttct atgagcgcct cgggtttacc gtcacagcgg
     4261 atgttgaagt ccctgaggga ccgcgcacat ggtgtatgac ccgaaagccc ggtgcctgat
     4321 tcgaaggcct gtcgtgaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4381 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4441 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4501 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4561 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     4621 tgctggctgc catgaactag ctactcggga ccccttaccg aaacatcgcc gcattctgca
     4681 gaggagtcga gtgtatgtaa acttctgacc cactgggaat gtgatgaaag aaataaaagc
     4741 tgaaatgaat cattctctct actattattc tgatatttca cattcttaaa ataaagtggt
     4801 gatcctaact gacctaagac agggaatttt tactaggatt aaatgtcagg aattgtgaaa
     4861 aagtgagttt aaatgtattt ggctaaggtg tatgtaaact tccgacttca actgtatagg
     4921 gatccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5521 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6841 aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6901 aggaaattgt aagcgttaat attttgttaa aattggatcc ctatacagtt gaagtcggaa
     6961 gtttacatac acttaagttg gagtcattaa aactcgtttt tcaactactc cacaaatttc
     7021 ttgttaacaa acaatagttt tggcaagtca gttaggacat ctactttgtg catgacacaa
     7081 gtcatttttc caacaattgt ttacagacag attatttcac ttataattca ctgtatcaca
     7141 attccagtgg gtcagaagtt tacatacact aagttcgact cctctgcaga atgcggcgat
     7201 gtttcggtaa ggggtccgct atctagtagg cccagctggt tctttccgcc tcagaagcca
     7261 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     7321 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     7381 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     7441 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag
     7501 ctctagagaa ttgatcccca agcttggcct gacaggcctt cgaattagtg agaaggaaag
     7561 acgcaaccag gaagcccacc aagagaaaga ggcatttcct gaaaagattc ccctttttgg
     7621 agagccctac aaggtattta ctgaacacta gacattgaag tcctgattta tcacaatgtt
     7681 gaaccctatg atgaaacaat tcagtataat tgagttcgaa actcgagcca tggtggcctc
     7741 ag
//
