LOCUS       sequence-97517-e        5157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding murine Pitx2 downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5157)
  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 5157)
  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-Pitx2
FEATURES             Location/Qualifiers
     source          1..5157
                     /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(1889..1913)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1986..2003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1992..2103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2477..2511
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2537..2553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2537..2553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2550..2572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2561..2577
                     /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(2585..2591,2592..2609,2610..2615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2630..2651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2768..2785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2939..3527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3019..3038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3698..4489,4490..4558))
                     /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     4321..4340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4559..4663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4731..4749
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4787..4809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4909..4928
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5122..5144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5136..5153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5137..5153
                     /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 tgaaggaatt cggtaccatg
      901 aactgcatga aaggcccgct gcccttggag caccgagcag ccgggactaa gctgtcggcc
      961 gcctcctcac ccttctgtca ccatccccag gcgttagcca tggcttcggt cctagctcct
     1021 ggccagcccc gctccttgga ctcctccaaa catagactgg aggtgcatac aatctccgat
     1081 acttccagcc ctgaagtcgc agagaaagat aagggccagc aaggaaagaa tgaggatgtg
     1141 ggcgccgagg acccgtccaa gaagaagcgg caacgccggc agaggactca tttcactagc
     1201 cagcagctgc aggagctgga agccactttc cagagaaacc gctacccaga catgtccact
     1261 cgcgaagaaa tcgccgtgtg gaccaacctt acggaagccc gagtccgggt ttggttcaag
     1321 aatcgccggg ccaaatggag aaagcgggaa cgcaaccagc aggccgagct gtgcaagaat
     1381 ggctttgggc cgcagttcaa cgggctcatg cagccctacg atgacatgta ccccggctat
     1441 tcgtacaaca attgggctgc caagggcctc acgtcagcgt ctctgtccac caagagcttc
     1501 cccttcttca actccatgaa cgtcaatccc ctgtcctctc agagtatgtt ttccccgccc
     1561 aactccatct catctatgag tatgtcgtcc agcatggtgc cctccgcggt gaccggcgtc
     1621 ccgggctcca gcctcaatag cctgaataac ttgaacaacc tgagcagccc gtcgctgaat
     1681 tccgcggtgc ccacgcccgc ctgtccttac gcgccgccga ctcctccgta cgtttatagg
     1741 gacacatgta actcgagcct ggccagcctg agactgaaag caaagcagca ctccagcttc
     1801 ggctacgcca gcgtgcagaa cccggcctcc aacctgagtg cttgccagta tgcagtcgac
     1861 cggccggtgt agctcgagtg cggccgcaac ccagctttct tgtacaaagt ggtgatccaa
     1921 ccggtataga tctatggccg gccatcaatt gatactagtc accgactcta gagctcgctg
     1981 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2041 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2101 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2161 gggggaggat tgggaagaca atagcaggca ctagtcggtc acgatatcta taacaagaaa
     2221 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
     2281 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
     2341 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
     2401 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
     2461 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaaaaaaa
     2521 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2581 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2641 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2701 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3361 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4681 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4741 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4801 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4861 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4921 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4981 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5041 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     5101 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaat
//
