LOCUS       sequence-223796-        8100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  AUTHORS   Lam PY, Sublett JE, Hollenbach AD, Roussel MF
  TITLE     The oncogenic potential of the Pax3-FKHR fusion protein requires the
            Pax3 homeodomain recognition helix but not the Pax3 paired-box DNA 
            binding domain.
  JOURNAL   Mol Cell Biol. 1999 Jan;19(1):594-601.
  PUBMED    9858583
REFERENCE   2  (bases 1 to 8100)
  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: "Mol Cell 
            Biol."; date: "1999-01"; volume: "19(1)"; pages: "594-601"
COMMENT     Sequence Label: pCDNA3-PAX3-FOXO1/FKHR
FEATURES             Location/Qualifiers
     source          1..8100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        552..755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     800..819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        800..818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3590..3608)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3591..3608)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3628..3645)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3634..3858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3904..4332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3991..4010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4201..4222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4341..4361)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4346..4675
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4526..4661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4588..4607
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4742..5536
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4796..4815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5406..5425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5710..5831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5747..5766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5801..5820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5880..5896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5880..5896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5893..5915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5904..5920
                     /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(5928..5934,5935..5952,5953..5958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5973..5994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6111..6128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6282..6870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6362..6381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7041..7832,7833..7901))
                     /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     7664..7683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7902..8006)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8081..8100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      781 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag
      841 ctcggatcct gcactcaagg gactcctctg gagcctgtgg acttggatct attagcgccc
      901 ccctccccgc gccccgcctc cctctctggc cttttttggg ggaggagctc tccgagatcc
      961 ggagagttcc cgagggtcac ccgccgcact gtgctcgctt tttcgtctcg ccttcacctg
     1021 gatataattt gcgagcgaag ctgcccccag gatgaccacg ctggccggcg ctgtgcccag
     1081 gatgatgcgg cccggcccgg ggcagaatta cccacgcagc ggcttcccgc tggaagtgtc
     1141 cacccctctt ggccagggcc gagtcaacca gctcggagga gtatttatca acggcaggcc
     1201 tctgcccaac catatccgcc acaagatagt ggagatggcc caccatggca ttcggccttg
     1261 cgtcatttct cgccagcttc gcgtgtccca tggttgcgtc tctaagatcc tgtgcaggta
     1321 ccaggagaca ggctccatcc gacctggtgc catcggcggc agcaaaccca agcaggtgac
     1381 aacgcctgac gtggagaaga aaattgagga atacaaaaga gagaacccgg gcatgtttag
     1441 ctgggaaatc agagacaaat tgctcaagga cgcggtctgt gatcgaaaca ccgtgccgtc
     1501 agtgagttcc atcagccgca tcctgagaag taaattcggg aaaggtgaag aggaggaggc
     1561 cgacttggag aggaaggagg cagaggaaag cgagaagaag gccaaacaca gcatcgacgg
     1621 catcctgagc gagcgagcct cagcacccca atcagatgaa ggctctgata ttgactctga
     1681 accagattta ccactaaaga ggaaacagcg cagaagccga accaccttca cagcagaaca
     1741 gctggaggaa ctggagcgtg cttttgagag aactcattac cctgacattt atactaggga
     1801 ggaactggcc cagagggcga agctcaccga ggcccgagta caggtctggt ttagcaaccg
     1861 ccgtgcaaga tggaggaagc aagctggggc caatcaactg atggctttca accatctcat
     1921 tcccgggggg ttccctccca ctgccatgcc gaccttgcca acgtaccagc tgtcggagac
     1981 ctcttaccag cccacatcta ttccacaagc tgtgtcagat cccagcagca ccgttcacag
     2041 acctcaaccg cttcctccaa gcactgtaca ccaaagcacg attccttcca acccagacag
     2101 cagctctgcc tactgcctcc ccagcaccag gcatggattt tccagctata cagacagctt
     2161 tgtgcctccg tcggggccct ccaaccccat gaaccccacc attggcaatg gcctctcacc
     2221 tcagaattca attcgtcata atctgtccct acacagcaag ttcattcgtg tgcagaatga
     2281 aggaactgga aaaagttctt ggtggatgct caatccagag ggtggcaaga gcgggaaatc
     2341 tcctaggaga agagctgcat ccatggacaa caacagtaaa tttgctaaga gccgaagccg
     2401 agctgccaag aagaaagcat ctctccagtc tggccaggag ggtgctgggg acagccctgg
     2461 atcacagttt tccaaatggc ctgcaagccc tggctctcac agcaatgatg actttgataa
     2521 ctggagtaca tttcgccctc gaactagctc aaatgctagt actattagtg ggagactctc
     2581 acccattatg accgaacagg atgatcttgg agaaggggat gtgcattcta tggtgtaccc
     2641 gccatctgcc gcaaagatgg cctctacttt acccagtctg tctgagataa gcaatcccga
     2701 aaacatggaa aatcttttgg ataatctcaa ccttctctca tcaccaacat cattaactgt
     2761 ttcgacccag tcctcacctg gcaccatgat gcagcagacg ccgtgctact cgtttgcgcc
     2821 accaaacacc agtttgaatt cacccagccc aaactaccaa aaatatacat atggccaatc
     2881 cagcatgagc cctttgcccc agatgcctat acaaacactt caggacaata agtcgagtta
     2941 tggaggtatg agtcagtata actgtgcgcc tggactcttg aaggagttgc tgacttctga
     3001 ctctcctccc cataatgaca ttatgacacc agttgatcct ggggtagccc agcccaacag
     3061 ccgggttctg ggccagaacg tcatgatggg ccctaattcg gtcatgtcaa cctatggcag
     3121 ccaggcatct cataacaaaa tgatgaatcc cagctcccat acccaccctg gacatgctca
     3181 gcagacatct gcagttaacg ggcgtcccct gccccacacg gtaagcacca tgccccacac
     3241 ctcgggtatg aaccgcctga cccaagtgaa gacacctgta caagtgcctc tgccccaccc
     3301 catgcagatg agtgccctgg ggggctactc ctccgtgagc agctgcaatg gctatggcag
     3361 aatgggcctt ctccaccagg agaagctccc aagtgacttg gatggcatgt tcattgagcg
     3421 cttagactgt gacatggaat ccatcattcg gaatgacctc atggatggag atacattgga
     3481 ttttaacttt gacaatgtgt tgcccaacca aagcttccca cacagtgtca agacaacgac
     3541 acatagctgg gtgtcaggct gactcgagca tgcatctaga gggccctatt ctatagtgtc
     3601 acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3661 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3721 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3781 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3841 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3901 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3961 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4021 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4081 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4141 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4201 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4261 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4321 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4381 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4501 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4561 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4621 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4681 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4741 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4801 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4861 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4921 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4981 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5041 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5101 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5161 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5221 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5281 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5341 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5401 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5461 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5521 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5581 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5641 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5701 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5761 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5821 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5881 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5941 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6001 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6061 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6181 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6961 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7021 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7081 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7141 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7201 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7261 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7321 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7381 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7441 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7501 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7561 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7621 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7681 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7741 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7801 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7861 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7921 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7981 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8041 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
//
