LOCUS       sequence-97516-e        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding murine Mycn downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  AUTHORS   Vereide DT, Vickerman V, Swanson SA, Chu LF, McIntosh BE, Thomson JA
  TITLE     An expandable, inducible hemangioblast state regulated by fibroblast
            growth factor.
  JOURNAL   Stem Cell Reports. 2014 Dec 9;3(6):1043-57. doi: 
            10.1016/j.stemcr.2014.10.003. Epub 2014 Nov 13.
  PUBMED    25458896
REFERENCE   2  (bases 1 to 5571)
  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"; doi: 
            "10.1016/j.stemcr.2014.10.003"; journalName: "Stem Cell Reports"; 
            date: "2014-12-9- 9"; volume: "3"; issue: "6"; pages: "1043-57"
COMMENT     Sequence Label: PB-Mycn
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..81
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    453..471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    490..508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    527..545
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    564..582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    638..656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    675..693
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    712..730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     752..776
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    856..880
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    1684..1695
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(2303..2327)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2400..2417)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2406..2517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2891..2925
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2951..2967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2951..2967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2964..2986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2975..2991
                     /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(2999..3005,3006..3023,3024..3029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3044..3065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3182..3199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3353..3941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3433..3452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4112..4903,4904..4972))
                     /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     4735..4754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4973..5077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5145..5163
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5201..5223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5323..5342
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5536..5558
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5550..5567
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5551..5567
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcaagcttta ttatttaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
       61 catgcgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
      121 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
      181 aatttattta tgtttattta tttattaaaa aaaaacaaaa actcaaaatt tcttctataa
      241 agtaacaaaa cttttaaaca ttctctcttt tacaaaaata aacttatttt gtactttaaa
      301 aacagtcatg ttgtaatcga taataagtaa ttagcttaac ctatacataa tagaaacaaa
      361 ttatacttat tagtcagtca gaaacaactt tggcacatat caatattatg ctctcgttaa
      421 ttaactcgag ggatccaagc tggccgcacg tctccctatc agtgatagag aagtcgacac
      481 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
      541 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
      601 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
      661 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
      721 gtgatagaga aggtaccccc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      781 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      841 gcggtggcgg ccatcacaag tttgtacaaa aaagcaggct tgaaggaatt cggtaccatg
      901 cccagctgca ccgcgtccac catgccgggg atgatctgca agaacccaga cctcgagttt
      961 gactcgctgc agccctgctt ctacccggac gaagatgact tctacttcgg cggtcccgac
     1021 tcgaccccac cgggggagga catctggaag aagtttgagc tgctgcccac gcccccgttg
     1081 tcgcccagcc gcgccttccc agagcacagc ccggagccct cgaattgggc tacggagatg
     1141 ctgctgccgg aggccgacct gtggggcaac ccggccgagg aggatgcgtt cggtctgggg
     1201 ggcctgggtg gcctcactcc taatccggtc atccttcagg actgcatgtg gagcggcttc
     1261 tctgcccgcg agaagctaga gcgcgcagtg aacgaaaaac tacagcacgg ccacgggccc
     1321 ccgggcgtca gctcagcctg ctcggctccc ggagtgggtg ccagcagccc cgggggccgt
     1381 gcccttggtg ggtcgtcgag tgctagccac accggggcca ccctgcctac cgacctctcc
     1441 cacccggctg ccgaatgtgt ggaccccgcc gtggtcttcc ccttcccggt gaacaagcga
     1501 gagtcggcgt cggtgcccgc tgcccccact agcgccccgg cgaccagcgc tgcggtcact
     1561 agtgtgtctg ttccagctac tgccccggtg gctgctcctg ctcgtgcagg cggccgtcct
     1621 gccagcagtg gggaggccaa ggccctcagc acctccggag aggatacctt gagcgactca
     1681 gatgatgagg atgacgagga ggaagatgaa gaggaggaaa tcgatgtggt caccgtagag
     1741 aagagacgtt cctcctctaa caacaaggcg gtaaccactt tcacgatcac tgtgcgtccc
     1801 aagacgtccg ctctgggcct ggggcgagca cagcctggcg agctgatcct caagcgctgt
     1861 gttcccatcc atcagcagca caactatgct gcaccctcac cctacgtgga gagcgaggac
     1921 gcgcccccgc agaaaaagat caagagcgag gcttctccac gccccctcaa aagtgttgtt
     1981 ccagcaaaag cgaagagcct gagcccccga aactcagact cggaggacag cgagcgccgc
     2041 cgcaaccaca acatcctgga gcgtcagcgc cggaacgacc tgcgctccag cttcctgacg
     2101 ctcagggacc atgtgcctga gctggtgaag aacgagaagg ccgccaaggt ggtcatcttg
     2161 aaaaaggcca ccgagtacgt gcacgcccta caggccaacg agcaccagct cctgctggaa
     2221 aaggagaaac tgcaggcgag gcagcagcag ttgctaaaga agatcgaaca cgctcggact
     2281 tgctagctcg agtgcggccg caacccagct ttcttgtaca aagtggtgat ccaaccggta
     2341 tagatctatg gccggccatc aattgatact agtcaccgac tctagagctc gctgatcagc
     2401 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2461 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2521 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2581 ggattgggaa gacaatagca ggcactagtc ggtcacgata tctataacaa gaaaatatat
     2641 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     2701 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     2761 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     2821 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     2881 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaaa aaaagcttgg
     2941 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3001 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3061 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3121 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3181 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3241 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3301 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3361 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3421 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3481 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3541 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3601 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3661 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3721 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3781 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3841 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3901 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3961 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4021 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4081 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4141 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4201 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4261 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4321 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4381 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4441 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4501 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4561 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4621 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4681 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4741 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4801 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4861 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4921 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4981 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5041 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5101 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5161 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5221 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5281 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     5341 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5401 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5461 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     5521 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa t
//
