LOCUS       sequence-268584-        7446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression of smURFP-NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7446)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 7446)
  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"
COMMENT     Sequence Label: BII-ChPtW-smURFP
FEATURES             Location/Qualifiers
     source          1..7446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..35
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(304..320)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(304..320)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1871..1924
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1931..1957
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1931..1957
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1964..2362
                     /codon_start=1
                     /product="dimeric small ultra-red fluorescent protein 
                     derived from a cyanobacterial phycobiliprotein (Rodriguez 
                     et al., 2016)"
                     /label=smURFP
                     /translation="MAKTSEQRVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAF
                     SQRDRALCLRDYGWFLHLITFCLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRC
                     LKEASLSLLDEEDANETAPYFDYIIKAMS"
     CDS             2366..2386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2418..3006
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2471..2491)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2889..2900)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3086..3220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3110..3129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3164..3183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3999..4061
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4191..4208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4191..4207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4349..4804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4436..4455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4646..4667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4830..4934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4935..5003,5004..5795)
                     /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(5153..5172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5966..6554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6455..6474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6708..6725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6842..6863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6878..6883,6884..6901,6902..6908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6916..6932
                     /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     6921..6943
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6940..6956
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6940..6956
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt
       61 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc
      121 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt
      181 gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa
      241 ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt
      301 atagatccac tagttctaga gggacagccc ccccccaaag cccccaggga tgtaattacg
      361 tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc
      421 cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg
      481 ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat
      541 acggggaaaa ggcctccaag gcctgcagga gtcaatggga aaaacccatt ggagccaagt
      601 acactgactc aatagggact ttccattggg ttttgcccag tacataaggt caataggggg
      661 tgagtcaaca ggaaagtccc attggagcca agtacattga gtcaataggg actttccaat
      721 gggttttgcc cagtacataa ggtcaatggg aggtaagcca atgggttttt cccattactg
      781 acatgtatac tgagtcatta gggactttcc aatgggtttt gcccagtaca taaggtcaat
      841 aggggtgaat caacaggaaa gtcccattgg agccaagtac actgagtcaa tagggacttt
      901 ccattgggtt ttgcccagta caaaaggtca atagggggtg agtcaatggg tttttcccat
      961 tattggcaca tacataaggt caataggggt gactagtgga gaagagcatg cttgagggct
     1021 gagtgcccct cagtgggcag agagcacatg gcccacagtc cctgagaagt tggggggagg
     1081 ggtgggcaat tgaactggtg cctagagaag gtggggcttg ggtaaactgg gaaagtgatg
     1141 tggtgtactg gctccacctt tttccccagg gtgggggaga accatatata agtgcagtag
     1201 tctctgtgaa cattcggttt agtgaacctg caggtcggcc gccacgaccg gtgccaccat
     1261 gactgaatac aaaccaactg ttcgcctggc aactcgtgat gatgttccac gtgcagttcg
     1321 caccctggct gctgcatttg ctgactaccc tgcaacccgt cacactgtgg acccagaccg
     1381 ccacattgaa cgtgtgactg aactgcagga gctgttcctg acccgtgtgg gcctggacat
     1441 tggcaaagtg tgggtggcag atgatggtgc tgctgtggca gtgtggacca cccctgaatc
     1501 tgttgaagct ggtgcagtgt ttgctgagat tggcccacgc atggcagaac tgtctggcag
     1561 ccgcctggca gcacaacagc agatggaagg tctgctggca ccacaccgcc caaaagaacc
     1621 tgcttggttc ctggcaactg tgggtgtgag ccctgaccac cagggtaagg gcctgggctc
     1681 tgcagtggtg ctgcctggtg tggaagcagc tgaacgtgca ggtgtgcctg ctttcctgga
     1741 gacctcagct ccacgcaacc tgcctttcta tgaacgcctg ggcttcactg tgactgctga
     1801 tgtggaagtg ccagaaggcc cacgcacttg gtgcatgact cgcaaaccag gtgctagcct
     1861 tggctccggc gagggcaggg gaagtcttct aacatgcggg gacgtggagg aaaatcccgg
     1921 cccactcggc tacccctacg acgtgcccga ctacgccagc ggcatggcta agacttccga
     1981 acagagggtg aacattgcta cactgctgac agaaaataag aagaaaatcg tggataaggc
     2041 ttcccaggat ctgtggcgga gacacccaga cctgatcgca ccaggaggaa ttgctttctc
     2101 tcagagggac cgcgctctgt gcctgcgaga ttacggctgg ttcctgcatc tgatcacctt
     2161 ttgtctgctg gccggagata agggccccat cgagtctatt gggctgatca gtattcgaga
     2221 aatgtataac tcactgggag tgcccgtccc tgcaatgatg gagagcatta gatgcctgaa
     2281 agaagccagc ctgtccctgc tggacgaaga ggacgccaac gagaccgcac cctactttga
     2341 ttacattatt aaggctatgt ccagccccaa gaagaagcgc aaggtgggcg cctaaggcta
     2401 gagttcgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2461 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2521 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2581 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2641 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2701 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2761 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     2821 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     2881 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2941 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3001 ccccgcctgg aattaattct ggctaagcac ttcgtggccg tcgatcgttt aaagggaggt
     3061 agtgagtcga gatatcgata ctagtgatcc agacatgata agatacattg atgagtttgg
     3121 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3181 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3241 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3301 caaatgtggt atggctgatt atgatcctct agagggacag ccccccccca aagcccccag
     3361 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     3421 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     3481 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     3541 cacctggggg gatacgggga aaaagccata ccaatggagc tcacggggac agcccccccc
     3601 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
     3661 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
     3721 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
     3781 tgagcctgca gacacctggg gggatacggg gaaaaagaat tcgataaaag ttttgttact
     3841 ttatagaaga aattttgagt ttttgttttt tttaataaat aaataaacat aaataaattg
     3901 tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa
     3961 attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacacatga
     4021 ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaatctag ctgcgtgttc
     4081 tgcagcgtgt cgagcatctt catctgctcc atcacgctgt aaaacacatt tgcaccgcga
     4141 gtctgcccgt cctccacggg ttcaaaaacg tgaatgaacg aggcgcgctc actggccgtc
     4201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4321 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4381 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4441 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4501 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4561 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4621 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4681 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4741 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4801 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4861 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4921 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4981 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5041 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5101 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5161 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5221 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5281 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5341 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5401 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5461 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5521 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5581 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5641 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5701 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5761 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5821 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5881 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5941 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6061 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6601 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6661 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6721 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6781 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6841 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6901 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6961 attacgccaa gcgcgcccgc cgggtaactc acggggtatc catgtccatt tctgcggcat
     7021 ccagccagga tacccgtcct cgctgacgta atatcccagc gccgcaccgc tgtcattaat
     7081 ctgcacaccg gcacggcagt tccggctgtc gccggtattg ttcgggttgc tgatgcgctt
     7141 cgggctgacc atccggaact gtgtccggaa aagccgcgac gaactggtat cccaggtggc
     7201 ctgaacgaac agttcaccgt taaaggcgtg catggccaca ccttcccgaa tcatcatggt
     7261 aaacgtgcgt tttcgctcaa cgtcaatgca gcagcagtca tcctcggcaa actctttcca
     7321 tgccgcttca acctcgcggg aaaaggcacg ggcttcttcc tccccgatgc ccagatagcg
     7381 ccagcttggg cgatgactga gccggaaaaa agacccgacg atatgatcct gatgcagcta
     7441 gattaa
//
