LOCUS       sequence-162385-        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"
     protein_bind    complement(96..120)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(200..224)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    246..264
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    283..301
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    320..338
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    357..375
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    431..449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    468..486
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    505..523
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     repeat_region   895..957
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(980..997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(980..996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(989..1011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1205..1224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1324..1346
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1384..1402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1470..1574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1575..1643,1644..2435)
                     /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(1793..1812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2606..3194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3095..3114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3348..3365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3482..3503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3518..3523,3524..3541,3542..3548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3556..3572
                     /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)."
     primer_bind     3561..3583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3580..3596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3580..3596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   3622..3656
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    4030..4141
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4130..4147
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    4220..4244
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4852..4863
                     /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)"
ORIGIN
        1 gggctgcagc gagtcaaact cgaggtctgg gttcttgcag atcatccccg gcatggtgga
       61 cgcggtgcag ctgggcatgg taccgaattc cttcaagcct gcttttttgt acaaacttgt
      121 gatggccgcc accgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc
      181 gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atataggggg
      241 taccttctct atcactgata gggagctcga gacgtgtcga cttctctatc actgataggg
      301 agacgttcta gacgtacctt ctctatcact gatagggagc tcgagacgtg tcgacttctc
      361 tatcactgat agggagacgt tctagacgta ccttctctat cactgatagg gagctcgaga
      421 cgtgtcgact tctctatcac tgatagggag acgttctaga cgtaccttct ctatcactga
      481 tagggagctc gagacgtgtc gacttctcta tcactgatag ggagacgtgc ggccagcttg
      541 gatccctcga gttaattaac gagagcataa tattgatatg tgccaaagtt gtttctgact
      601 gactaataag tataatttgt ttctattatg tataggttaa gctaattact tattatcgat
      661 tacaacatga ctgtttttaa agtacaaaat aagtttattt ttgtaaaaga gagaatgttt
      721 aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttta ataaataaat
      781 aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa
      841 cttaatatct attcaaatta ataaataaac ctcgatatac agaccgataa aacacatgcg
      901 tcaattttac gcatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta
      961 aataataaag cttgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1021 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1081 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     1141 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     1201 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     1261 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     1321 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     1381 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1441 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1561 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1681 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1741 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1801 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1861 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1981 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2101 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2161 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2281 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2401 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3361 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3421 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3481 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3541 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     3601 attacgccaa gcttttttta accctagaaa gatagtctgc gtaaaattga cgcatgcatt
     3661 cttgaaatat tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat
     3721 ctcagtcgcc gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat
     3781 tttgaactat aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt
     3841 aagatttaac tcatacgata attatattgt tatttcatgt tctacttacg tgataactta
     3901 ttatatatat attttcttgt tatagatatc gtgaccgact agtgcctgct attgtcttcc
     3961 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     4021 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     4081 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     4141 gtcgaggctg atcagcgagc tctagagtcg gtgactagta tcaattgatg gccggccata
     4201 gatctatacc ggttggatca ccactttgta caagaaagct gggttgcggc cgcactcgag
     4261 ctagcaagtc cgagcgtgtt cgatcttctt tagcaactgc tgctgcctcg cctgcagttt
     4321 ctccttttcc agcaggagct ggtgctcgtt ggcctgtagg gcgtgcacgt actcggtggc
     4381 ctttttcaag atgaccacct tggcggcctt ctcgttcttc accagctcag gcacatggtc
     4441 cctgagcgtc aggaagctgg agcgcaggtc gttccggcgc tgacgctcca ggatgttgtg
     4501 gttgcggcgg cgctcgctgt cctccgagtc tgagtttcgg gggctcaggc tcttcgcttt
     4561 tgctggaaca acacttttga gggggcgtgg agaagcctcg ctcttgatct ttttctgcgg
     4621 gggcgcgtcc tcgctctcca cgtagggtga gggtgcagca tagttgtgct gctgatggat
     4681 gggaacacag cgcttgagga tcagctcgcc aggctgtgct cgccccaggc ccagagcgga
     4741 cgtcttggga cgcacagtga tcgtgaaagt ggttaccgcc ttgttgttag aggaggaacg
     4801 tctcttctct acggtgacca catcgatttc ctcctcttca tcttcctcct cgtcatcctc
     4861 atcatctgag tcgctcaagg tatcctctcc ggaggtgctg agggccttgg cctccccact
     4921 gctggcagga cggccgcctg cacgagcagg agcagccacc ggggcagtag ctggaacaga
     4981 cacactagtg accgcagcgc tggtcgccgg ggcgctagtg ggggcagcgg gcaccgacgc
     5041 cgactctcgc ttgttcaccg ggaaggggaa gaccacggcg gggtccacac attcggcagc
     5101 cgggtgggag aggtcggtag gcagggtggc cccggtgtgg ctagcactcg acgacccacc
     5161 aagggcacgg cccccggggc tgctggcacc cactccggga gccgagcagg ctgagctgac
     5221 gcccgggggc ccgtggccgt gctgtagttt ttcgttcact gcgcgctcta gcttctcgcg
     5281 ggcagagaag ccgctccaca tgcagtcctg aaggatgacc ggattaggag tgaggccacc
     5341 caggcccccc agaccgaacg catcctcctc ggccgggttg ccccacaggt cggcctccgg
     5401 cagcagcatc tccgtagccc aattcgaggg ctccgggctg tgctctggga aggcgcggct
     5461 gggcgacaac gggggcgtgg gcagcagctc aaacttcttc cagatgtcct cccccggtgg
     5521 ggtcgagtcg ggaccgccga agtagaagtc atcttcgtcc gggtagaagc a
//
