LOCUS       sequence-200842-        6579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector encoding doxycycline-inducible human MYCN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   Miller AZ, Satchie A, Tannenbaum AP, Nihal A, Thomson JA, Vereide DT
  TITLE     Expandable Arterial Endothelial Precursors from Human CD34(+) Cells 
            Differ in Their Proclivity to Undergo an Endothelial-to-Mesenchymal 
            Transition.
  JOURNAL   Stem Cell Reports. 2018 Jan 9;10(1):73-86. doi: 
            10.1016/j.stemcr.2017.12.011.
  PUBMED    29320761
REFERENCE   2  (bases 1 to 6579)
  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.2017.12"; journalName: "Stem Cell Reports"; date: 
            "2018-01-9- 9"; volume: "10"; issue: "1"; pages: "73-86"
COMMENT     Sequence Label: PB-TRE3G-MYCN
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..53)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     153..172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     366..388
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     380..397
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   420..482
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1430..1808
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1438..1456
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1474..1492
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1510..1528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1546..1564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1582..1600
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1618..1636
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1654..1672
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1717..1741
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1826..1850
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3283..3307)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3357..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3368..3387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3422..3441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3949..3982)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     repeat_region   4300..4334
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4360..4376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4360..4376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4373..4395)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4384..4400
                     /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(4408..4414,4415..4432,4433..4438))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4453..4474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4591..4608)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4762..5350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4842..4861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5521..6312,6313..6381))
                     /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     6144..6163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6382..6486)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6554..6572
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
       61 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcaagcttt attatttaac
      421 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
      481 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
      541 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
      601 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttaaac
      661 attctctctt ttacaaaaat aaacttattt tgtactttaa aaacagtcat gttgtaatcg
      721 ataataagta attagcttaa cctatacata atagaaacaa attatactta ttagtcagtc
      781 agaaacaact ttggcacata tcaatattat gctctcgtta attaactcga gggatccggc
      841 gcgcctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
      901 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
      961 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     1021 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     1081 cctgggggct ttgggagggg gctgtccccg tgagctccca ggcgcgcctg tcattctaaa
     1141 tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca ggctcaaaga
     1201 gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc
     1261 cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc
     1321 ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg
     1381 ggggctgtcc ccgtgagctc ccaggcgcgc cgctagccaa gaattcctcg agtttactcc
     1441 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
     1501 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
     1561 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
     1621 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtataa
     1681 gctttaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga accgtcagat
     1741 cgcctggagc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
     1801 ctcgtaaacc gcggtggcgg ccatcacaag tttgtacaaa aaagcaggct taaggagttt
     1861 aaacaccatg ccgagctgct ccacgtccac catgccgggc atgatctgca agaacccaga
     1921 cctcgagttt gactcgctac agccctgctt ctacccggac gaagatgact tctacttcgg
     1981 cggccccgac tcgacccccc cgggggagga catctggaag aagtttgagc tgctgcccac
     2041 gcccccgctg tcgcccagcc gtggcttcgc ggagcacagc tccgagcccc cgagctgggt
     2101 cacggagatg ctgcttgaga acgagctgtg gggcagcccg gccgaggagg acgcgttcgg
     2161 cctgggggga ctgggtggcc tcacccccaa cccggtcatc ctccaggact gcatgtggag
     2221 cggcttctcc gcccgcgaga agctggagcg cgccgtgagc gagaagctgc agcacggccg
     2281 cgggccgcca accgccggtt ccaccgccca gtccccggga gccggcgccg ccagccctgc
     2341 gggtcgcggg cacggcgggg ctgcgggagc cggccgcgcc ggggccgccc tgcccgccga
     2401 gctcgcccac ccggccgccg agtgcgtgga tcccgccgtg gtcttcccct ttcccgtgaa
     2461 caagcgcgag ccagcgcccg tgcccgcagc cccggccagt gccccggcgg cgggccctgc
     2521 ggtcgcctcg ggggcgggta ttgccgcccc agccggggcc ccgggggtcg cccctccgcg
     2581 cccaggcggc cgccagacca gcggcggcga ccacaaggcc ctcagtacct ccggagagga
     2641 caccctgagc gattcagatg atgaagatga tgaagaggaa gatgaagagg aagaaatcga
     2701 cgtggtcact gtggagaagc ggcgttcctc ctccaacacc aaggctgtca ccacattcac
     2761 catcactgtg cgtcccaaga acgcagccct gggtcccggg agggctcagt ccagcgagct
     2821 gatcctcaaa cgatgccttc ccatccacca gcagcacaac tatgccgccc cctctcccta
     2881 cgtggagagt gaggatgcac ccccacagaa gaagataaag agcgaggcgt ccccacgtcc
     2941 gctcaagagt gtcatccccc caaaggctaa gagcttgagc ccccgaaact ctgactcgga
     3001 ggacagtgag cgtcgcagaa accacaacat cctggagcgc cagcgccgca acgaccttcg
     3061 gtccagcttt ctcacgctca gggaccacgt gccggagttg gtaaagaatg agaaggccgc
     3121 caaggtggtc attttgaaaa aggccactga gtatgtccac tccctccagg ccgaggagca
     3181 ccagcttttg ctggaaaagg aaaaattgca ggcaagacag cagcagttgc taaagaaaat
     3241 tgaacacgct cggacttgct agaagagccc gggcgatcgc gaacccagct ttcttgtaca
     3301 aagtggtgat ccaaccggta tagatctcga gctcgatgag tttaaaccag acatgataag
     3361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3541 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatccggctg cctcgcgctt
     3601 aattaacccg ggggccgcct gggaactgcc gctggccccc caccgcccca aggatctccg
     3661 gtccccgccc ggcttgctga cgtcacggat gccccagggt gtgctgggca ggtagcggga
     3721 gcctgggaaa tggagtccat tagcagaagt ggccacttcc aggagtcgct gtgccccgat
     3781 gcacactggg aagtccgcag ctccgaggcc cccagtggaa atcgccagat gagggcctcc
     3841 tccggggaat gctgggaaat ggagtctaca ggccggaggg gtccccacgg catactagca
     3901 agtgtgtgta gcaccgcgta aaggggatga atagcagact gccccgggat aacttcgtat
     3961 aggatacttt atacgaagtt atgagctctg tgataatcac tagtgaattg atactagtcg
     4021 gtcacgatat ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg
     4081 aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg
     4141 cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca
     4201 agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat
     4261 tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga
     4321 ctatctttct agggttaaaa aaagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4381 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4441 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4501 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4561 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4621 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4681 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4741 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4801 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4861 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4921 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4981 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5041 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5101 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5161 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5221 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5281 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5341 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5401 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5461 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5521 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5581 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5641 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5701 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5761 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5821 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5881 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5941 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6001 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6061 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6121 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6181 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6241 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6301 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6361 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6421 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6481 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6541 attaacctat aaaaataggc gtatcacgag gccctttcg
//
