LOCUS       sequence-348394-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty transposon-based vector for mammalian polycistronic 
            expression of moxGFP, c-Myc and an additional gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7814)
  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 7814)
  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-cMyc-Puro
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        181..285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(286..354,355..1146)
                     /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(504..523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1317..1905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1806..1825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2059..2076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2450..2469
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2468..2523
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2569..2588
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2688..3287)
                     /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(2771..2791)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3268..3287
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        3919..5101
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     5049..5069
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      5127..5136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5859..5915
                     /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             5937..5990
                     /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"
     CDS             6951..6977
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7221..7250
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7333..7350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7339..7563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agttgaaaaa cgagttttaa tgactccaac ttaagtgtat gtaaacttcc gacttcaact
       61 gtatagggat ccaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
      121 tccttacgca tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg
      181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      241 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      301 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      361 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      421 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      481 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      541 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      601 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      661 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      721 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      781 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      841 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      901 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      961 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1021 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1081 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1141 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1201 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1261 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1321 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1381 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1441 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1501 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1561 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1621 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1681 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1741 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1801 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1861 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1921 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1981 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2041 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggatcccta tacagttgaa
     2101 gtcggaagtt tacatacacc ttagccaaat acatttaaac tcactttttc acaattcctg
     2161 acatttaatc ctagtaaaaa ttccctgtct taggtcagtt aggatcacca ctttatttta
     2221 agaatgtgaa atatcagaat aatagtagag agaatgattc atttcagctt ttatttcttt
     2281 catcacattc ccagtgggtc agaagtttac atacactcga ctcctctgca gaatgcggcg
     2341 atgtttcggt aaggggtccc gagtagctag ttcatggcag ccagcatatg gcatatgttg
     2401 ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc
     2461 ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt
     2521 attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg
     2581 gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct
     2641 gataggcagc ctgcacctga ggagtgaatt cacgacaggc cttcgaatca ggcaccgggc
     2701 ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg
     2761 aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc
     2821 accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg
     2881 ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg
     2941 tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg
     3001 gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc
     3061 cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg
     3121 cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg
     3181 gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg
     3241 acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt ggcctcaggt
     3301 gcagaggttt ctacagggaa agagagaaag gctacagtta gtttgctgtg atgatgctaa
     3361 gtgccgaggt cagatgtcta tggattgtac acggaatccc aacgcaggaa ccaaaaaaca
     3421 ctcacccaaa aaaatggcgc taaggccgag agaaaggcga agttccgtct acggctatat
     3481 accttgaaac aggggcggca gaagtgacgc aacgcggttg cgcggacggc ttcctgtgtt
     3541 ttcattggtc gaagcgttct gcgcgtgcgc agagattccc cagggaaggc gggccctctt
     3601 ccgaatgtcc aatccggaca ttcttccggt tggacctccg cggtcagcgc tgtgcccggc
     3661 gagagatcac gtggggcgcg gaggcggtgc tgctggggca cggccgtcca gcctcggcgg
     3721 ccatattttt gaggggctgt tcatctcgtt cacacgctct gtccgccatg tttgtgagtg
     3781 gaagcgccat taccccttca agcgactgaa ggctgcaggg cctctggtgg cccgcatggg
     3841 gagaccagac ccgccaggcc cgcctttccg cactcagtcc gggcttactt tattttgtga
     3901 gacagggtct cgcctagagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     3961 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     4021 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     4081 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     4141 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
     4201 ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga
     4261 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     4321 tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa tctggtggca
     4381 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     4441 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     4501 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     4561 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     4621 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     4681 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     4741 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     4801 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     4861 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     4921 gggggagggg ttttatgcga tggagtttcc ccatactgag tgggtggaga ctgaagttag
     4981 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     5041 gttcattctc aagcctcaga cagtggttca aagttttttt cttccattta aggtgtcgtg
     5101 aaaactaccc caagctggcc tctgaggcca ccatggtgtc aaagggggag gaactgttta
     5161 ctggggtcgt gcctattctg gtcgaactgg atggagatgt caacggccac aaattcagcg
     5221 tgaggggaga gggagagggc gacgcaacca atggcaagct gacactgaag ttcatctcta
     5281 ccacaggcaa gctgcccgtg ccttggccaa ccctggtgac cacactgaca tacggcgtgc
     5341 agagcttttc caggtatcct gaccacatga agagacacga tttctttaag tccgccatgc
     5401 cagagggcta cgtgcaggag cggaccatct ctttcaagga cgatggcacc tataagacac
     5461 gcgccgaggt gaagtttgag ggcgacacac tggtgaacag gatcgagctg aagggcatcg
     5521 acttcaagga ggatggcaat atcctgggcc acaagctgga gtacaacttt aatagccaca
     5581 acgtgtatat caccgccgat aagcagaaga acggcatcaa ggccaatttc aagatcagac
     5641 acaatgtgga ggacggctcc gtgcagctgg ccgatcacta ccagcagaac accccaatcg
     5701 gcgacggacc agtgctgctg cccgataatc actatctgtc tacacagagc aagctgtcca
     5761 aggaccccaa cgagaagcgg gatcacatgg tgctgctgga gttcgtgacc gcagcaggaa
     5821 tcacacacgg aatggacgag ctgtacaagg gctctggcgc caccaacttt agcctgctga
     5881 agcaggcagg cgatgtggag gagaatcccg gccctgatat cgtcgacggc agcggcgagg
     5941 gaaggggctc actgctgact tgcggcgatg tggaggagaa cccaggaccc gggatgcccc
     6001 tgaacgtgag cttcaccaac aggaattacg acctggatta tgactccgtg cagccttact
     6061 tctattgcga tgaggaggag aacttctacc agcagcagca gcagagcgag ctgcagccac
     6121 ctgcaccttc cgaggacatc tggaagaagt ttgagctgct gcctacccca cccctgtctc
     6181 caagcaggag atctggcctg tgctccccct cttatgtggc cgtgacacct ttcagcctga
     6241 gaggcgataa cgacggagga ggaggaagct tttccaccgc cgatcagctg gagatggtga
     6301 cagagctgct gggcggcgat atggtgaacc agtccttcat ctgcgaccct gacgatgaga
     6361 cctttatcaa gaatatcatc atccaggatt gtatgtggtc tggcttcagc gccgcagcaa
     6421 agctggtgtc cgagaagctg gcctcttacc aggcagcaag gaaggactct ggaagcccta
     6481 acccagccag gggccactcc gtgtgctcta caagctccct gtacctgcag gatctgtctg
     6541 ccgccgccag cgagtgtatc gacccatccg tggtgttccc ctatcctctg aatgattcta
     6601 gctcccccaa gagctgcgcc tcccaggatt ctagcgcctt tagcccttcc tctgactccc
     6661 tgctgagctc caccgagtct agcccacagg gaagcccaga gccactggtg ctgcacgagg
     6721 agacacctcc aaccacatcc tctgactccg aggaggagca ggaggatgag gaggagatcg
     6781 acgtggtgtc tgtggagaag aggcaggccc ccggcaagag gagcgagtct ggaagcccat
     6841 ccgccggagg acactctaag ccacctcaca gcccactggt gctgaagcgg tgccacgtgt
     6901 ccacccacca gcacaactac gccgccccac cctctacaag gaaggattat ccagccgcca
     6961 agagagtgaa gctggactcc gtgagggtgc tgagacagat ctccaacaat aggaagtgta
     7021 cctctcccag aagctccgat acagaggaga acgtgaagcg gcgcacccac aatgtgctgg
     7081 agcgccagag gagaaatgag ctgaagcgga gcttctttgc cctgcgcgac cagatccctg
     7141 agctggagaa caatgagaag gccccaaagg tggtcatcct gaagaaggcc acagcctata
     7201 tcctgtctgt gcaggccgag gagcagaagc tgatcagcga ggaggacctg ctgaggaaga
     7261 ggagggagca gctgaagcac aagctggagc agctgagaaa tagctgtgcc tgatctagag
     7321 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     7381 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     7441 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     7501 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     7561 tggcttctga ggcggaaaga accagctggg cctactagat agcggacccc ttaccgaaac
     7621 atcgccgcat tctgcagagg agtcgaactt agtgtatgta aacttctgac ccactggaat
     7681 tgtgatacag tgaattataa gtgaaataat ctgtctgtaa acaattgttg gaaaaatgac
     7741 ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca aaactattgt ttgttaacaa
     7801 gaaatttgtg gagt
//
