LOCUS       sequence-97464-e        5622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding rat Gata2 downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  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 5622)
  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-Gata2
FEATURES             Location/Qualifiers
     source          1..5622
                     /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    complement(2354..2378)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2451..2468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2457..2568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2942..2976
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3002..3018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3002..3018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3015..3037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3026..3042
                     /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(3050..3056,3057..3074,3075..3080))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3095..3116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3233..3250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3404..3992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3484..3503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4163..4954,4955..5023))
                     /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     4786..4805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5024..5128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5196..5214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5252..5274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5374..5393
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5587..5609
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5601..5618
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5602..5618
                     /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 ccgaattcgc ccttaggatg
      901 gaggtggcgc ctgagcagcc tcgctggatg gcgcaccccg ccgtattgaa tgcgcagcac
      961 cccgactcgc accatccggg cctggcacac aactacatgg agccggcaca gctgctgcct
     1021 ccggacgagg tggatgtctt cttcaaccac ctcgactcgc agggcaaccc ctactacgcc
     1081 aacccagccc acgcgcgcgc gcgcgtttcc tacagtccag cgcatgcccg tctcacagga
     1141 ggccagatgt gccgaccaca cttgttgcac agcccaggct tgccatggct ggacgggggc
     1201 aaagcagctc tctctgccgc tgctgcccat caccacagcc cctggaccgt cagcccgttc
     1261 tccaagaccc cgctgcaccc ctcagctgct ggagcacctg gagggcctct gtctgtgtac
     1321 ccaggggctg cgggtgggag cggtggaggc agcgggagct ccgtagcctc cctcaccccc
     1381 actgcagccc actcaggctc ccatctcttc ggcttcccac ccacgccacc caaagaagtg
     1441 tctccagacc ccagcaccac aggagctgct tccccagcct caccttcagc agggggtagt
     1501 gtagcccggg gtgaggacaa ggatggcgtc aagtatcaag tgtcactgtc tgagagcatg
     1561 aagatggaag gtggcagtcc cctgcgcccg ggcctagctc ccatgggcac ccagcctgca
     1621 acacaccacc caatacccac ctatccctcc tatgtgcctg cctcagctca tgaatatggc
     1681 agcgggctct tccatcccgg aggcttcctg ggtggcccag cctccagctt cacccctaag
     1741 cagcgaagca aggctcgctc ctgctcagaa ggccgggagt gtgtcaactg cggggccaca
     1801 gccacccctc tctggcgacg ggatggcacg ggccactacc tgtgcaatgc ctgtgggctc
     1861 taccacaaga tgaatggaca gaatcggccg ctcatcaagc ccaagaggag actgtctgct
     1921 gccaggagag cgggcacctg ttgtgcaaat tgtcagacga caaccaccac cttatggcgc
     1981 cggaacgcca acggggaccc tgtgtgcaac gcctgtggcc tctactacaa gctgcacaat
     2041 gttaacaggc cactgaccat gaagaaggaa gggatccaga cccggaatcg gaagatgtcc
     2101 agcaaatcca agaagagcaa gaaaggagcc gaatgctttg aggagctatc caagtgcatg
     2161 caggagaagt catccccctt cagtgcggct gccctggctg gacacatggc acctgtggga
     2221 cacctcccac cttttagtca ctctggacac atcctaccta cgccaacgcc tatccacccc
     2281 tcctccagtc tctcctttgg tcacccccac ccgtccagca tggtgactgc catgggctag
     2341 aagggcgaat tcgacccagc tttcttgtac aaagtggtga tccaaccggt atagatctat
     2401 ggccggccat caattgatac tagtcaccga ctctagagct cgctgatcag cctcgactgt
     2461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2641 agacaatagc aggcactagt cggtcacgat atctataaca agaaaatata tatataataa
     2701 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     2761 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     2821 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
     2881 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     2941 gcatgcgtca attttacgca gactatcttt ctagggttaa aaaaagcttg gcgtaatcat
     3001 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3061 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3121 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3181 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3241 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3301 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3361 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3421 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3481 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3541 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3601 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3661 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3721 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3781 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3841 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3901 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4081 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4141 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4201 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4261 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4321 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4381 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4441 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4501 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4561 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4621 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4681 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4741 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4801 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4861 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4921 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4981 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5041 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5161 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5221 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5281 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5341 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5401 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5461 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5521 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5581 ggttttccca gtcacgacgt tgtaaaacga cggccagtga at
//
