LOCUS       sequence-209732-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty transposon containing SV-40-driven 
            Venus-LEQLE-SIINFEKL-TEW for antigen presentation studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  TITLE     SIINFEKL antigen presentation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5057)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pT2/SV-Venus-SIINFEKL
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..56
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     48..65
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     49..65
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..94
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(488..508)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        493..822
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      673..808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(834..836,837..839,840..1550)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(879..900)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1140..1159)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1487..1508
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1761..1895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1798..1817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1852..1871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2318..2338)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2318..2336)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2357..2373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2357..2373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2370..2392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2381..2397
                     /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(2405..2411,2412..2429,2430..2435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2450..2471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2588..2605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2759..3347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2839..2858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3518..4309,4310..4378))
                     /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     4141..4160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4379..4483)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4510..4965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4597..4616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4807..4828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
       61 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctcgga tccctataca
      121 gttgaagtcg gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta
      181 ctccacaaat ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt
      241 gtgcatgaca caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat
      301 tcactgtatc acaattccag tgggtcagaa gtttacatac actaagttga ctgtgccttt
      361 aaacagcttg gaaaattcca gaaaatgatg tcatggcttt agaagcttcc cacaataagt
      421 tgggtgaatt ttggctcatt cctcctttct ataggattga ggtcagagct ttgtgatgga
      481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
      541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
      781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggatggtga
      841 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      901 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      961 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1021 ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1081 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1141 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1201 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1261 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1321 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1381 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1441 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1501 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ctcgagcagc
     1561 tggaatccat catcaacttc gaaaagctca ctgaatggtg attcgaaatg accgaccaag
     1621 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1681 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1741 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     1801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1861 ccaaactcat caatgtatct tatcatgtct ggatcccttg aaatacatcc acaggtacac
     1921 ctccaattga ctcaaatgat gtcaattagt ctatcagaag cttgtggaag gctactcgaa
     1981 atgtttgacc caagttaaac aatttaaagg caatgctacc aaatactaat tgagtgtatg
     2041 taaacttctg acccactggg aatgtgatga aagaaataaa agctgaaatg aatcattctc
     2101 tctactatta ttctgatatt tcacattctt aaaataaagt ggtgatccta actgacctaa
     2161 gacagggaat ttttactagg attaaatgtc aggaattgtg aaaaagtgag tttaaatgta
     2221 tttggctaag gtgtatgtaa acttccgact tcaactgtat agggatcctc tagctagagt
     2281 cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     2341 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2401 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2461 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2941 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3181 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4381 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4441 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4501 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4561 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4621 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4681 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4741 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4801 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4861 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4921 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag
     4981 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5041 gaaaggggga tgtgctg
//
