LOCUS       sequence-268580-        7483 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Piggybac vector for ASCL2 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 7483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: BII-STAR-tdT
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      473..482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             479..1906
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             1910..1930
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1959..1976)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1965..2189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2873..2935
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3065..3082)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3065..3081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3074..3096)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3223..3678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3310..3329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3520..3541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3704..3808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3809..3877,3878..4669)
                     /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     complement(4027..4046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4840..5428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5329..5348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5582..5599
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5716..5737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5752..5757,5758..5775,5776..5782)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5790..5806
                     /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)."
     primer_bind     5795..5817
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5814..5830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5814..5830
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   6321..6355
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     misc_feature    join(6896..6961,6962..6999)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     primer_bind     complement(7019..7038)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7063..7111
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    7299..7347
                     /note="synthetic polyadenylation signal"
     misc_feature    7361..7452
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7401..7420
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcctatccc agcttctata ggtgaaaagg caggtgcaaa gcaaatccta tcccagcttc
       61 tataggtgaa aaggcaggtg caaagcaaat cctatcccag cttctatagg tgaaaaggca
      121 ggtgcaaagc aaatcctatc ccagcttcta taggtgaaaa ggcaggtgca aagcaaatcc
      181 tatcccagct tctataggtg aaaaggcagg tgcaaagcaa atcctatccc agcttctact
      241 aggtgaaaag gcaggtgcaa agcaaatcct atcccagctt ctatagggag gggacaagtg
      301 gagggaaagt ggggccgggc ggggggtgcc tgggaagcca ggctgcgctg acgtcactgg
      361 gcgcgcaatt cgggctggag cgctttaaaa aacgagcgtg caagcagaga tgctgctcca
      421 caccgctcag gccgcgaccg gagcttggca ttccggtact gttgccaact cggccaccat
      481 ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
      541 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      601 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
      661 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
      721 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      781 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
      841 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
      901 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
      961 gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat
     1021 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
     1081 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     1141 ccaccacctg ttcctggggc atggcaccgg cagcaccggc agcggcagct ccggcaccgc
     1201 ctcctccgag gacaacaaca tggccgtcat caaagagttc atgcgcttca aggtgcgcat
     1261 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1321 cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     1381 ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga
     1441 catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1501 cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct
     1561 gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa
     1621 gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa
     1681 gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa
     1741 gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa
     1801 gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga
     1861 gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagggcc ccaagaagaa
     1921 gcgcaaggtg ggcgcctaag ctaaaaaccg ctgatcagcc tcgactgtgc cttctagttg
     1981 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2041 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2101 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2161 gcatgctggg gatgcggtgg gctctatggc ctgttaggcc ttctagaggg acagcccccc
     2221 cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc
     2281 ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga
     2341 cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc
     2401 tttgagcctg cagacacctg gggggatacg gggaaaaagc cataccaatg gagctcacgg
     2461 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
     2521 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
     2581 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
     2641 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gaattcgata
     2701 aaagttttgt tactttatag aagaaatttt gagtttttgt tttttttaat aaataaataa
     2761 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
     2821 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
     2881 aattttacac atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaat
     2941 ctagctgcgt gttctgcagc gtgtcgagca tcttcatctg ctccatcacg ctgtaaaaca
     3001 catttgcacc gcgagtctgc ccgtcctcca cgggttcaaa aacgtgaatg aacgaggcgc
     3061 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     3121 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3181 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3241 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3301 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3361 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3421 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3481 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3541 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3601 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3661 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3781 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3841 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3901 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3961 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4021 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4081 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4141 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4201 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4261 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4321 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4381 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4441 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4501 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4561 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4621 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4681 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4741 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4801 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4861 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4921 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5461 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5521 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5581 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5641 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5701 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5761 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5821 cagctatgac catgattacg ccaagcgcgc ccgccgggta actcacgggg tatccatgtc
     5881 catttctgcg gcatccagcc aggatacccg tcctcgctga cgtaatatcc cagcgccgca
     5941 ccgctgtcat taatctgcac accggcacgg cagttccggc tgtcgccggt attgttcggg
     6001 ttgctgatgc gcttcgggct gaccatccgg aactgtgtcc ggaaaagccg cgacgaactg
     6061 gtatcccagg tggcctgaac gaacagttca ccgttaaagg cgtgcatggc cacaccttcc
     6121 cgaatcatca tggtaaacgt gcgttttcgc tcaacgtcaa tgcagcagca gtcatcctcg
     6181 gcaaactctt tccatgccgc ttcaacctcg cgggaaaagg cacgggcttc ttcctccccg
     6241 atgcccagat agcgccagct tgggcgatga ctgagccgga aaaaagaccc gacgatatga
     6301 tcctgatgca gctagattaa ccctagaaag atagtctgcg taaaattgac gcatgcattc
     6361 ttgaaatatt gctctctctt tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc
     6421 tcagtcgccg cttggagctc ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt
     6481 ttgaactata acgaccgcgt gagtcaaaat gacgcatgat tatcttttac gtgactttta
     6541 agatttaact catacgataa ttatattgtt atttcatgtt ctacttacgt gataacttat
     6601 tatatatata ttttcttgtt atagatctgt ggaatgtgtg ctagagggac agcccccccc
     6661 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
     6721 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
     6781 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
     6841 tgagcctgca gacacctggg gggatacggg gaaaaggcct ggtaggctgc agcggtaggg
     6901 cgcagtagtc cagggtttcc ttgatgatgt catacttatc ctgtcccttt tttttccaca
     6961 gctcgcggtt gaggacaaac tcttcgcggt ctttccagta tgcatggggg aggctaactg
     7021 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     7081 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     7141 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     7201 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     7261 gcaggccctg ccatagcata ttcggccgcc acggccgcaa taaaatatct ttattttcat
     7321 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     7381 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     7441 gaacatttct ctatcgatag gtgaaaaggc aggtgcaaag caa
//
