LOCUS       sequence-394152-        7364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown of FYN tyrosine kinase. Construct has inverted repeats to 
            be used in Sleeping Beauty transposon system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7364)
  AUTHORS   Comba A, Dunn PJ, Argento AE, Kadiyala P, Ventosa M, Patel P, Zamler
            DB, Nunez FJ, Zhao L, Castro MG, Lowenstein PR
  TITLE     Fyn tyrosine kinase, a downstream target of receptor tyrosine 
            kinases, modulates antiglioma immune responses.
  JOURNAL   Neuro Oncol. 2020 Jun 9;22(6):806-818. doi: 10.1093/neuonc/noaa006.
  PUBMED    31950181
REFERENCE   2  (bases 1 to 7364)
  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"; doi: "10"; journalName: 
            "Neuro Oncol"; date: "2020-06-9- 9"; volume: "22"; issue: "6"; 
            pages: "806-818"
COMMENT     Sequence Label: pT2/sh-FYN1-GFP_Seq_C
FEATURES             Location/Qualifiers
     source          1..7364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             1034..1550
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1614..1955
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2022..2438
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2368..2390
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2406..2422
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           2478..2571
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2618..2632
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2660..2788
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(2821..2841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2826..3155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3006..3141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3162..3164,3165..3167,3168..3881)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3207..3228)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3468..3487)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3815..3836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3954..4468
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4630..4646
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4654..4660,4661..4678,4679..4684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4699..4720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5179..5199)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5179..5197)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5218..5234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5218..5234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5242..5258
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5266..5272,5273..5290,5291..5296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5311..5332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5449..5466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5620..6208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5700..5719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6379..7170,7171..7239))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7002..7021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7240..7344)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg agtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct
      661 cggatcccta tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac
      721 tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt
      781 aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt
      841 atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag
      901 ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc
      961 ttgatctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     1021 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     1081 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     1141 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     1201 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     1261 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     1321 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     1381 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     1441 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
     1501 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
     1561 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
     1621 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1681 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     1741 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     1801 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     1861 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     1921 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     1981 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     2041 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     2101 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     2161 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     2221 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     2281 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     2341 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     2401 cctcccttta tccagccctc actccttctc taggcgccgg aattagatct ctcgactagg
     2461 gataacaggg taattgtttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat
     2521 cttggaaaca cttgctggga ttacttcttc aggttaaccc aacagaaggc tcgagtgctg
     2581 ttgacagtga gcgcccacac taacttcctg tataatagtg aagccacaga tgtattatac
     2641 aggaagttag tgtggttgcc tactgcctcg gagaattcaa ggggctactt taggagcaat
     2701 tatcttgttt actaaaactg aataccttgc tatctctttg atacattttt acaaagctga
     2761 attaaaatgg tataaattaa atcacttttt tcaattcgcc agcacagtgg tcgaccctgt
     2821 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2881 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3001 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3061 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3121 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct catggtgagc aagggcgagg
     3181 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3241 agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3301 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3361 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3421 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3481 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3541 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3601 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gcgaacttca
     3661 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3721 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3781 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3841 ccgccgggat cactctcggc atggacgagc tgtacaagta atgaattaat taagaattat
     3901 caagcttatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     3961 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     4021 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     4081 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4141 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     4201 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4261 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4321 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct
     4381 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4441 tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca
     4501 ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc
     4561 cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga
     4621 attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4741 cgctttccag tcgggaaacc tgtcgtgcca gatccatgga cttcactagt gcgcgcggcc
     4801 gctctagatg gccagatcta gcttgtggaa ggctactcga aatgtttgac ccaagttaaa
     4861 caatttaaag gcaatgctac caaatactaa ttgagtgtat gtaaacttct gacccactgg
     4921 gaatgtgatg aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat
     4981 ttcacattct taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag
     5041 gattaaatgt caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta
     5101 aacttccgac ttcaactgta tagggatcct ctagctagag tcgacctcgc gggggggccc
     5161 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt
     6841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgc
//
