LOCUS       sequence-97515-e        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding murine Lmo2 downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  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 4866)
  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-Lmo2
FEATURES             Location/Qualifiers
     source          1..4866
                     /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(1598..1622)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1695..1712)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1701..1812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2186..2220
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2246..2262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2246..2262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2259..2281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2270..2286
                     /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(2294..2300,2301..2318,2319..2324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2339..2360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2477..2494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2648..3236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2728..2747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3407..4198,4199..4267))
                     /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     4030..4049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4268..4372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4440..4458
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4496..4518)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4618..4637
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4831..4853
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4845..4862
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4846..4862
                     /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 attcgctctc tctctttggc gtttgaaggg agcgcggtga ctgtccttga gcgtggaggg
      961 gcgagctcgc cagcggagcg accgagcaag aggaggcgca ggagcggcgg cgcccgagca
     1021 cccgaggggg tccgagtcca ggcagctagc cagccacgcg ccacaaaggg agcgcccccg
     1081 ccgcccggca cgccgccccc ctccccaatg tcctcggcca tcgaaaggaa gagcctggac
     1141 ccgtctgagg aacccgtgga tgaggtgctg cagatacccc catccctgct gacatgtggt
     1201 ggctgccagc agaacatagg ggaccgctac ttcctgaaag ccatcgacca gtactggcat
     1261 gaggattgcc tcagctgtga cctctgtggg tgtcggctgg gagaggtggg gaggcgcctc
     1321 tactacaagc tgggacggaa attgtgcagg agagactatc tcaggctttt tggtcaggat
     1381 ggtctctgtg catcctgtga caagcggatc cgtgcctatg agatgacgat gcgggtgaaa
     1441 gacaaagtgt atcacctgga gtgtttcaaa tgcgccgcct gtcagaagca tttctgtgta
     1501 ggtgacagat accttctcat caactccgac atagtgtgtg aacaagacat ctacgagtgg
     1561 accaagatca atgggatcat ctagaagggc gaattcgacc cagctttctt gtacaaagtg
     1621 gtgatccaac cggtatagat ctatggccgg ccatcaattg atactagtca ccgactctag
     1681 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1741 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1801 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1861 ggacagcaag ggggaggatt gggaagacaa tagcaggcac tagtcggtca cgatatctat
     1921 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
     1981 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
     2041 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
     2101 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
     2161 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
     2221 ttaaaaaaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2281 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2341 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2401 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2461 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2521 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2581 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2641 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2701 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2761 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2821 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2881 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2941 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3001 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3061 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3121 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3181 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3241 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3421 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3481 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3541 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3601 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4321 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4381 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4441 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4501 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4561 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4621 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4681 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     4741 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4801 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4861 gtgaat
//
