LOCUS       sequence-97519-e        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding murine Tal1 downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  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 5169)
  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-Tal1
FEATURES             Location/Qualifiers
     source          1..5169
                     /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(1901..1925)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1998..2015)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2004..2115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2489..2523
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2549..2565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2549..2565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2562..2584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2573..2589
                     /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(2597..2603,2604..2621,2622..2627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2642..2663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2780..2797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2951..3539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3031..3050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3710..4501,4502..4570))
                     /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     4333..4352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4571..4675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4743..4761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4799..4821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4921..4940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5134..5156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5148..5165
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5149..5165
                     /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 acggagcggc cgccgagcga ggcggcacgc agtgacccgc aactagaggg acaggacgcg
      961 gccgaggccc gcatggcccc cccgcaccta gtcctgctca acggcgtcgc caaggagacg
     1021 agccgcgcag ccccggctga gccccccgtc atcgagctag gagcgcgcag cggcgcgggg
     1081 ggcggccctg ccagtggggg cggtgccgcg agggacttaa agggccgcga cgcagtagca
     1141 gccgaagctc gccttcgggt gcccaccacc gagctgtgca gacctcccgg acccgccccg
     1201 gcgcccgcgc ccgcctcggc tcctgcagag ctgcctggag acggccgcat ggtgcagctg
     1261 agcccgcccg cgctggcagc ccctgccggc cccggccgag cgctgctcta tagccttagc
     1321 cagccgctcg cctcactagg cagtgggttc tttggggaac cggatgcctt ccccatgttc
     1381 accaacaaca accgggtgaa gaggaggccc tccccatatg agatggagat ttctgatggt
     1441 cctcacacca aagtagtgcg gcgcatcttc accaacagcc gggaacgatg gcggcagcag
     1501 aatgtgaatg gggcatttgc tgagctcaga aagctgatcc ccacccaccc accagacaag
     1561 aaactaagca agaatgagat cctccgcctt gccatgaagt acatcaattt cctggccaag
     1621 ttactcaatg accaggagga ggaaggcacc cagcgtgcca agcctggcaa ggaccccgtg
     1681 gtgggagctg gtgggggtgg ggcagggggt ggcatccccc ctgaagacct tctacaggac
     1741 gtgctttccc ccaactccag ctgtggcagc tctctggatg gggcagccag cccggacagt
     1801 tacacagagg agccaacacc caagcacact tcccgcagcc tccatcctgc cctgctgcct
     1861 gccgctgatg gggctggccc ccggtgaaag ggcgaattcg acccagcttt cttgtacaaa
     1921 gtggtgatcc aaccggtata gatctatggc cggccatcaa ttgatactag tcaccgactc
     1981 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2041 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2101 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2161 gcaggacagc aagggggagg attgggaaga caatagcagg cactagtcgg tcacgatatc
     2221 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     2281 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     2341 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     2401 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     2461 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     2521 gggttaaaaa aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2581 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2641 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2701 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2761 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2821 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2881 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2941 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3001 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3061 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3121 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3181 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3241 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3301 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3361 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3421 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3481 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3541 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3601 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3661 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3721 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3781 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3841 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3901 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3961 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4021 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4081 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4141 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4201 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4261 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4321 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4381 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4441 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4501 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4561 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4621 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4681 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4741 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4801 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4861 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4921 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     4981 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     5041 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5101 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5161 ccagtgaat
//
