LOCUS       sequence-383136-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Consitiutive mammalian expression of membrane bound human IL-21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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 G418R mbIL21
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1268..1285)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1274..1498
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1930..2034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2035..2103,2104..2895)
                     /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(2253..2272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3066..3654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3555..3574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3808..3825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4199..4218
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4217..4272
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4318..4337
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(4412..5206)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4523..4542)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5133..5152
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        5857..6068
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     6968..6988
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ctgaggccac catgtggctt caatctctgt tgctcttggg cacggttgcg tgcagtatca
       61 gtcaagggca agatagacac atgattcgga tgcggcagtt gattgatatt gtggaccaac
      121 tcaaaaacta tgtcaatgat ctcgtccctg agtttctccc tgcgccggaa gacgttgaaa
      181 ctaattgtga gtggagtgcc ttttcttgct ttcaaaaggc tcagcttaag agtgccaata
      241 ctgggaacaa tgagcgaatc atcaatgtca gcatcaaaaa actgaagcga aagcctcctt
      301 caacgaatgc cggtcgaagg caaaagcacc ggttgacgtg tcccagttgt gatagctacg
      361 aaaagaagcc ccctaaagaa tttctcgaaa gattcaaaag tctgttgcaa aagatgatcc
      421 accagcatct cagttctcgg acgcatggca gtgaagactc cgagtccaaa tatgggcctc
      481 cgtgtccacc atgtcccgct ccagaatttt tgggaggacc tagcgtattt ctgtttcctc
      541 cgaaaccaaa ggatacgttg atgatttctc ggactccgga ggtgacctgt gtcgtagtcg
      601 atgtatcaca agaggacccc gaggtgcagt tcaattggta cgtcgatggc gtcgaagtac
      661 acaatgccaa gaccaagcct agagaggaac agtttaactc tacgtacagg gtagtatcag
      721 ttttgacagt gctgcatcag gattggctca acggcaagga gtacaaatgt aaagtttcca
      781 ataagggtct cccctcatcc attgagaaga ctatctcaaa ggctaaaggg cagcccagag
      841 aaccacaggt atatacactg ccccctagtc aggaagagat gacgaaaaat caagtatcct
      901 tgacgtgctt ggttaaagga ttctatccta gcgatattgc tgtagaatgg gagtctaatg
      961 ggcagcccga gaataattat aagaccactc ccccagtgtt ggattcagac gggtccttct
     1021 ttctttactc ccgccttacc gttgataaga gtcggtggca ggaagggaat gtattttcat
     1081 gttccgtgat gcacgaggca cttcataacc attacactca gaagtctttg agtttgagct
     1141 tgggcaagat ggcgttgatc gtacttggcg gtgtcgcggg tctgttgctt ttcatcggac
     1201 tgggaatatt tttctaaggc ctgtcaggcc aagcttgggg atcaattctc tagagctcgc
     1261 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1321 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1381 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1441 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1501 tctgaggcgg aaagaaccag ctgggcctac tagatagcgg accccttacc gaaacatcgc
     1561 cgcattctgc agaggagtcg aacttagtgt atgtaaactt ctgacccact ggaattgtga
     1621 tacagtgaat tataagtgaa ataatctgtc tgtaaacaat tgttggaaaa atgacttgtg
     1681 tcatgcacaa agtagatgtc ctaactgact tgccaaaact attgtttgtt aacaagaaat
     1741 ttgtggagta gttgaaaaac gagttttaat gactccaact taagtgtatg taaacttccg
     1801 acttcaactg tatagggatc caattttaac aaaatattaa cgcttacaat ttcctgatgc
     1861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catcaggtgg cacttttcgg
     1921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggatccctat
     3841 acagttgaag tcggaagttt acatacacct tagccaaata catttaaact cactttttca
     3901 caattcctga catttaatcc tagtaaaaat tccctgtctt aggtcagtta ggatcaccac
     3961 tttattttaa gaatgtgaaa tatcagaata atagtagaga gaatgattca tttcagcttt
     4021 tatttctttc atcacattcc cagtgggtca gaagtttaca tacactcgac tcctctgcag
     4081 aatgcggcga tgtttcggta aggggtcccg agtagctagt tcatggcagc cagcatatgg
     4141 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     4201 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     4261 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     4321 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     4381 ccaccttctg ataggcagcc tgcacctgag gtcagaagaa ctcgtcaaga aggcgataga
     4441 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     4501 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     4561 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     4621 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg
     4681 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     4741 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     4801 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     4861 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     4921 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     4981 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg
     5041 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     5101 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     5161 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatggtg gcctcaggtg
     5221 cagaggtttc tacagggaaa gagagaaagg ctacagttag tttgctgtga tgatgctaag
     5281 tgccgaggtc agatgtctat ggattgtaca cggaatccca acgcaggaac caaaaaacac
     5341 tcacccaaaa aaatggcgct aaggccgaga gaaaggcgaa gttccgtcta cggctatata
     5401 ccttgaaaca ggggcggcag aagtgacgca acgcggttgc gcggacggct tcctgtgttt
     5461 tcattggtcg aagcgttctg cgcgtgcgca gagattcccc agggaaggcg ggccctcttc
     5521 cgaatgtcca atccggacat tcttccggtt ggacctccgc ggtcagcgct gtgcccggcg
     5581 agagatcacg tggggcgcgg aggcggtgct gctggggcac ggccgtccag cctcggcggc
     5641 catatttttg aggggctgtt catctcgttc acacgctctg tccgccatgt ttgtgagtgg
     5701 aagcgccatt accccttcaa gcgactgaag gctgcagggc ctctggtggc ccgcatgggg
     5761 agaccagacc cgccaggccc gcctttccgc actcagtccg ggcttacttt attttgtgag
     5821 acagggtctc gcctagaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     5881 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     5941 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     6001 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     6061 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     6121 cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag
     6181 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
     6241 cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat ctggtggcac
     6301 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
     6361 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
     6421 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     6481 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     6541 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     6601 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     6661 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     6721 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     6781 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     6841 ggggaggggt tttatgcgat ggagtttccc catactgagt gggtggagac tgaagttagg
     6901 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     6961 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttaa ggtgtcgtga
     7021 aaactacccc aagctggcct
//
