LOCUS       sequence-483811-        8314 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Empty Sleeping Beauty XPRESSO vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8314)
  TITLE     XPRESSO: Rapid Genetic Engineering of Human Pluripotent Stem Cells 
            for Durable Overexpression Using a Modular Anti-silencing Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSB-XPRESSO
FEATURES             Location/Qualifiers
     source          1..8314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    324..548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     537..554
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    587..1175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             693..704
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1102..1122
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      1389..1398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1460..1479)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1487..2503
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(1541..1563)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        2782..3161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             4332..5033
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(4465..4483)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4733..4752
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4927..4950
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             5043..5099
                     /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             5100..5699
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5100..5119)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5596..5616
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5799..5818)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5864..5919
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5918..5937)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6311..6328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6482..7070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6562..6581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7241..8032,8033..8101))
                     /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     7864..7883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8102..8206)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca
      301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      541 aggcacagtc gaggctgatc agcgagctca ggtcgacggt atcgatgcgg ggaggcggcc
      601 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
      661 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
      721 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat
      781 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
      841 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
      901 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
      961 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     1021 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     1081 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     1141 aatctttcac aaattttgta atccagaggt tgattatcga aagcttggcc tgacaggcct
     1201 tcgaattagt gagaaggaaa gacgcaacca ggaagcccac caagagaaag aggcatttcc
     1261 tgaaaagatt cccctttttg gagagcccta caaggtattt actgaacact agacattgaa
     1321 gtcctgattt atcacaatgt tgaaccctat gatgaaacaa ttcagtataa ttgagttcga
     1381 aactcgagcc atggtggcct cagaggccaa cagcggccgc gatatcctcg aggaattctt
     1441 tgccaaaatg atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga
     1501 agaaggcatg aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga
     1561 accccgccct gccccgtccc ccccgaaggc agccgtcccc ccgcggacag ccccgaggct
     1621 ggagagggag aaggggacgg cggcgcggcg acgcacgaag gccctccccg cccatttcct
     1681 tcctgccggc gccgcaccgc ttcgccccgc gcccgctaga gggggtgcgg cggcgcctcc
     1741 cagatttcgg ctccgcacag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt
     1801 accataaaag gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccgggggc
     1861 cgccgcgccc ctcccccgag cccyccccgg cccgaggcgg ccccgccccg cccggcaccc
     1921 ccacctgccg ccaccccccg cccggcacgg cgagccccgc gccacgcccc gtacggagcc
     1981 ccgcacccga agccgggccg tgctcagcaa ctcggggagg ggggtgcagg gggggttgca
     2041 gcccgaccga cgcgcccaca ccccctgctc acccccccac gcacacaccc cgcacgcagc
     2101 ctttgttccc ctcgcagccc cccccgcacc gcggggcacc gcccccggcc gcgctcccct
     2161 cgcgcacact gcggagcgca caaagccccg cgccgcgccc gcagcgctca cagccgccgg
     2221 gcagcgcgga gccgcacgcg gcgctcccca cgcacacaca cacgcacgca ccccccgagc
     2281 cgctcccccc gcacaaaggg ccctcccgga gcccctcaag gctttcacgc agccacagaa
     2341 aagaaacaag ccgtcattaa accaagcgct aattacagcc cggaggagaa gggccgtccc
     2401 gcccgctcac ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg
     2461 cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac tcccgcccgc cgcgcgcttc
     2521 gctttttata gggccgccgc cgccgccgcc tcgccataaa aggaaacttt cggagcgcgc
     2581 cgctctgatt ggctgccgcc gcacctcycc scctcscccc sccccccccc tccccccccc
     2641 cccccccccc ctggcgcgcc cccccccccc ccccgccccc atcgctgcac aaaataatta
     2701 aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg gagatgggga gagtgaagca
     2761 gaacgtgggg ctcacctcga ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2821 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2881 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     2941 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     3001 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     3061 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     3121 cgtaattgat tactattaat aactagtcaa taatcaatgt cgactcgtat tcccgggctg
     3181 ccaagcagaa aagccaccgc tgaggagact ccggtcactg tcctcgcccc gcctccccct
     3241 tccctcccct tggggaccac cgggcgccac gccgcgaacg gtaagtgccg cggtcgtcgg
     3301 cgcctccgcc ctccccctag ggccccaatt cccagcgggc gcggcgccgg cccctccccc
     3361 cgccgcgcgc gcgcccgctg ccccgccctt cgtggccgcc cggcgtgggc ggtgccaccc
     3421 ctccccccgg cggccccgcg cgcagctccc ggctccctcc cccttcggat gtggcttgag
     3481 ctgtaggcgc ggagggccgg agacgctgca gacccgcgac ccggagcagc tcggaggcgg
     3541 tgaagtcggt ggctttcctt ctctctagct ctcgctcgct ggtggtgctt cagatgccac
     3601 acgcgtagct tataataccg gttcaattgc cgacccctcc ccccaacttc tcggggactg
     3661 tgggcgatgt gcgctctgcc cactgacggg caccggagcc tctaggcgag accctgtctc
     3721 acaaaataaa gtaagcccgg actgagtgcg gaaaggcggg cctggcgggt ctggtctccc
     3781 catgcgggcc accagaggcc ctgcagcctt cagtcgcttg aaggggtaat ggcgcttcca
     3841 ctcacaaaca tggcggacag agcgtgtgaa cgagatgaac agcccctcaa aaatatggcc
     3901 gccgaggctg gacggccgtg ccccagcagc accgcctccg cgccccacgt gatctctcgc
     3961 cgggcacagc gctgaccgcg gaggtccaac cggaagaatg tccggattgg acattcggaa
     4021 gagggcccgc cttccctggg gaatctctgc gcacgcgcag aacgcttcga ccaatgaaaa
     4081 cacaggaagc cgtccgcgca accgcgttgc gtcacttctg ccgcccctgt ttcaaggtat
     4141 atagccgtag acggaacttc gcctttctct cggccttagc gccatttttt tgggtgagtg
     4201 ttttttggtt cctgcgttgg gattccgtgt acaatccata gacatctgac ctcggcactt
     4261 agcatcatca cagcaaacta actgtagcct ttctctcttt ccctgtagaa acctctgcac
     4321 ctgaggccac catgttgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg
     4381 tgcgcatgga gggctccatg aacggccatg agttcgagat cgagggcgag ggcgagggcc
     4441 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     4501 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     4561 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     4621 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     4681 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     4741 tgcagaagaa gacaatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     4801 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
     4861 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     4921 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     4981 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagggcagtg
     5041 gagctactaa cttcagcctg ctgaagcagg ctggtgacgt cgaggagaat cctggcccca
     5101 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     5161 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5221 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5281 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5341 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5401 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5461 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5521 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5581 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5641 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat
     5701 tcgaaggcct gtcgtgaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     5761 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     5821 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     5881 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5941 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     6001 tgctggctgc catgaactag ctactcggga ccccttaccg aaacatcgcc gcattctgca
     6061 gaggagtcga gtgtatgtaa acttctgacc cactgggaat gtgatgaaag aaataaaagc
     6121 tgaaatgaat cattctctct actattattc tgatatttca cattcttaaa ataaagtggt
     6181 gatcctaact gacctaagac agggaatttt tactaggatt aaatgtcagg aattgtgaaa
     6241 aagtgagttt aaatgtattt ggctaaggtg tatgtaaact tccgacttca actgtatagg
     6301 gatccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8221 aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     8281 aggaaattgt aagcgttaat attttgttaa aatt
//
