LOCUS       sequence-348395-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty transposon-based vector for mammalian polycistronic 
            expression of moxGFP and two additional genes of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  AUTHORS   Acon M, Geiss C, Torres-Calvo J, Bravo-Estupinan D, Oviedo G, 
            Arias-Arias JL, Rojas-Matey LA, Edwin B, Vasquez-Vargas G, 
            Oses-Vargas Y, Guevara-Coto J, Segura-Castillo A, Siles-Canales F, 
            Quiros-Barrantes S, Regnier-Vigouroux A, Mendes P, Mora-Rodriguez R
  TITLE     MYC dosage compensation is mediated by miRNA-transcription factor 
            interactions in aneuploid cancer.
  JOURNAL   iScience. 2021 Nov 8;24(12):103407. doi: 10.1016/j.isci.2021.103407.
            eCollection 2021 Dec 17.
  PUBMED    34877484
REFERENCE   2  (bases 1 to 6497)
  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: "iScience.";
            date: "2021-11-8"; volume: "24(12):103407. doi"; pages: " 
            10.1016/j.isci.2021.103407. eCollection 2021 Dec 17"
COMMENT     Sequence Label: pSBbi-moxGFP-P2A-T2A-Puro
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(225..293,294..1085)
                     /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(443..462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1256..1844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1745..1764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1998..2015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2389..2408
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2407..2462
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2508..2527
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2627..3226)
                     /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(2710..2730)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3207..3226
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        3858..5040
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     4988..5008
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      5066..5075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5798..5854
                     /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             5876..5929
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(5955..5972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5961..6185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tatagggatc caattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
       61 ccttacgcat ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc
      121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      181 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      241 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      301 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      361 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      421 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
      481 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
      541 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
      601 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
      661 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
      721 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
      781 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
      841 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
      901 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
      961 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1021 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1081 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1141 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1201 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1261 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1321 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1381 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1441 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1501 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1561 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1621 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1681 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1741 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1801 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1861 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1921 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1981 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggatccctat acagttgaag
     2041 tcggaagttt acatacacct tagccaaata catttaaact cactttttca caattcctga
     2101 catttaatcc tagtaaaaat tccctgtctt aggtcagtta ggatcaccac tttattttaa
     2161 gaatgtgaaa tatcagaata atagtagaga gaatgattca tttcagcttt tatttctttc
     2221 atcacattcc cagtgggtca gaagtttaca tacactcgac tcctctgcag aatgcggcga
     2281 tgtttcggta aggggtcccg agtagctagt tcatggcagc cagcatatgg catatgttgc
     2341 caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc catatgtcct
     2401 tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa atttccttta
     2461 ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt tggcagaggg
     2521 aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca ccaccttctg
     2581 ataggcagcc tgcacctgag gagtgaattc acgacaggcc ttcgaatcag gcaccgggct
     2641 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
     2701 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca
     2761 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
     2821 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
     2881 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
     2941 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
     3001 agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
     3061 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg
     3121 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga
     3181 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtg gcctcaggtg
     3241 cagaggtttc tacagggaaa gagagaaagg ctacagttag tttgctgtga tgatgctaag
     3301 tgccgaggtc agatgtctat ggattgtaca cggaatccca acgcaggaac caaaaaacac
     3361 tcacccaaaa aaatggcgct aaggccgaga gaaaggcgaa gttccgtcta cggctatata
     3421 ccttgaaaca ggggcggcag aagtgacgca acgcggttgc gcggacggct tcctgtgttt
     3481 tcattggtcg aagcgttctg cgcgtgcgca gagattcccc agggaaggcg ggccctcttc
     3541 cgaatgtcca atccggacat tcttccggtt ggacctccgc ggtcagcgct gtgcccggcg
     3601 agagatcacg tggggcgcgg aggcggtgct gctggggcac ggccgtccag cctcggcggc
     3661 catatttttg aggggctgtt catctcgttc acacgctctg tccgccatgt ttgtgagtgg
     3721 aagcgccatt accccttcaa gcgactgaag gctgcagggc ctctggtggc ccgcatgggg
     3781 agaccagacc cgccaggccc gcctttccgc actcagtccg ggcttacttt attttgtgag
     3841 acagggtctc gcctagaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     3901 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     3961 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     4021 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     4081 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     4141 cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag
     4201 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
     4261 cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat ctggtggcac
     4321 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
     4381 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
     4441 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     4501 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     4561 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     4621 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     4681 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     4741 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     4801 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     4861 ggggaggggt tttatgcgat ggagtttccc catactgagt gggtggagac tgaagttagg
     4921 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     4981 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttaa ggtgtcgtga
     5041 aaactacccc aagctggcct ctgaggccac catggtgtca aagggggagg aactgtttac
     5101 tggggtcgtg cctattctgg tcgaactgga tggagatgtc aacggccaca aattcagcgt
     5161 gaggggagag ggagagggcg acgcaaccaa tggcaagctg acactgaagt tcatctctac
     5221 cacaggcaag ctgcccgtgc cttggccaac cctggtgacc acactgacat acggcgtgca
     5281 gagcttttcc aggtatcctg accacatgaa gagacacgat ttctttaagt ccgccatgcc
     5341 agagggctac gtgcaggagc ggaccatctc tttcaaggac gatggcacct ataagacacg
     5401 cgccgaggtg aagtttgagg gcgacacact ggtgaacagg atcgagctga agggcatcga
     5461 cttcaaggag gatggcaata tcctgggcca caagctggag tacaacttta atagccacaa
     5521 cgtgtatatc accgccgata agcagaagaa cggcatcaag gccaatttca agatcagaca
     5581 caatgtggag gacggctccg tgcagctggc cgatcactac cagcagaaca ccccaatcgg
     5641 cgacggacca gtgctgctgc ccgataatca ctatctgtct acacagagca agctgtccaa
     5701 ggaccccaac gagaagcggg atcacatggt gctgctggag ttcgtgaccg cagcaggaat
     5761 cacacacgga atggacgagc tgtacaaggg ctctggcgcc accaacttta gcctgctgaa
     5821 gcaggcaggc gatgtggagg agaatcccgg ccctgatatc gtcgacggca gcggcgaggg
     5881 aaggggctca ctgctgactt gcggcgatgt ggaggagaac ccaggacccg ggtaatctag
     5941 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6001 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     6061 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     6121 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     6181 tatggcttct gaggcggaaa gaaccagctg ggcctactag atagcggacc ccttaccgaa
     6241 acatcgccgc attctgcaga ggagtcgaac ttagtgtatg taaacttctg acccactgga
     6301 attgtgatac agtgaattat aagtgaaata atctgtctgt aaacaattgt tggaaaaatg
     6361 acttgtgtca tgcacaaagt agatgtccta actgacttgc caaaactatt gtttgttaac
     6421 aagaaatttg tggagtagtt gaaaaacgag ttttaatgac tccaacttaa gtgtatgtaa
     6481 acttccgact tcaactg
//
