LOCUS       sequence-419051-        5825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty Transposase expression vector driven by the CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  AUTHORS   Strom AR, Biggs RJ, Banigan EJ, Wang X, Chiu K, Herman C, Collado J,
            Yue F, Ritland Politz JC, Tait LJ, Scalzo D, Telling A, Groudine M, 
            Brangwynne CP, Marko JF, Stephens AD
  TITLE     HP1alpha is a chromatin crosslinker that controls nuclear and 
            mitotic chromosome mechanics.
  JOURNAL   Elife. 2021 Jun 9;10. pii: 63972. doi: 10.7554/eLife.63972.
  PUBMED    34106828
REFERENCE   2  (bases 1 to 5825)
  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: "Elife."; 
            date: "2021-06-9"; volume: "10. pii: 63972. doi"; pages: " 
            10.7554/eLife.63972"
COMMENT     Sequence Label: SB100X in pCAG globin pA
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1733..1742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1739..2761
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL
                     KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     primer_bind     complement(2847..2866)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2912..2967
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2966..2985)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3328..3344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3328..3344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3341..3363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3352..3368
                     /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(3376..3382,3383..3400,3401..3406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3421..3442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3501..3696
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3547..3682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3702..3836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3739..3758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3793..3812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3904..3921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4075..4663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4155..4174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4834..5625,5626..5694))
                     /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     5457..5476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5695..5799)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg ggggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgccy cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gggaaaatca aaagaaatca gccaagacct cagaaaaaga attgtagacc tccacaagtc
     1801 tggttcatcc ttgggagcaa tttccaaacg cctggcggta ccacgttcat ctgtacaaac
     1861 aatagtacgc aagtataaac accatgggac cacgcagccg tcataccgct caggaaggag
     1921 acgcgttctg tctcctagag atgaacgtac tttggtgcga aaagtgcaaa tcaatcccag
     1981 aacaacagca aaggaccttg tgaagatgct ggaggaaaca ggtacaaaag tatctatatc
     2041 cacagtaaaa cgagtcctat atcgacataa cctgaaaggc cactcagcaa ggaagaagcc
     2101 actgctccaa aaccgacata agaaagccag actacggttt gcaactgcac atggggacaa
     2161 agatcgtact ttttggagaa atgtcctctg gtctgatgaa acaaaaatag aactgtttgg
     2221 ccataatgac catcgttatg tttggaggaa gaagggggag gcttgcaagc cgaagaacac
     2281 catcccaacc gtgaagcacg ggggtggcag catcatgttg tgggggtgct ttgctgcagg
     2341 agggactggt gcacttcaca aaatagatgg catcatggac gcggtgcagt atgtggatat
     2401 attgaagcaa catctcaaga catcagtcag gaagttaaag cttggtcgca aatgggtctt
     2461 ccaacacgac aatgacccca agcatacttc caaagttgtg gcaaaatggc ttaaggacaa
     2521 caaagtcaag gtattggagt ggccatcaca aagccctgac ctcaatccta tagaaaattt
     2581 gtgggcagaa ctgaaaaagc gtgtgcgagc aaggaggcct acaaacctga ctcagttaca
     2641 ccagctctgt caggaggaat gggccaaaat tcacccaaat tattgtggga agcttgtgga
     2701 aggctacccg aaacgtttga cccaagttaa acaatttaaa ggcaatgcta ccaaatacta
     2761 ggcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     2821 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     2881 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     2941 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3001 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     3061 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     3121 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     3181 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     3241 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3301 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3481 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3541 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3601 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3661 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     3721 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3781 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     3841 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5821 acctg
//
