LOCUS       sequence-256381-        7445 bp    DNA     circular SYN 15-APR-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 7445)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 7445)
  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-ChPtW-smURFP
FEATURES             Location/Qualifiers
     source          1..7445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1875..1928
                     /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             1935..1961
                     /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             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1968..2366
                     /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             2370..2390
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2422..3010
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2475..2495)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2893..2904)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3090..3224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3114..3133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3168..3187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4002..4064
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4194..4211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4194..4210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4203..4225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4352..4807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4439..4458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4649..4670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4833..4937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4938..5006,5007..5798)
                     /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(5156..5175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6711..6728
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6845..6866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6881..6886,6887..6904,6905..6911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6919..6935
                     /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     6924..6946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6943..6959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6943..6959
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      601 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      661 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
      721 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
      781 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
      841 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
      901 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
      961 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1021 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1081 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1141 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1201 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1261 ccatgactga atacaaacca actgttcgcc tggcaactcg tgatgatgtt ccacgtgcag
     1321 ttcgcaccct ggctgctgca tttgctgact accctgcaac ccgtcacact gtggacccag
     1381 accgccacat tgaacgtgtg actgaactgc aggagctgtt cctgacccgt gtgggcctgg
     1441 acattggcaa agtgtgggtg gcagatgatg gtgctgctgt ggcagtgtgg accacccctg
     1501 aatctgttga agctggtgca gtgtttgctg agattggccc acgcatggca gaactgtctg
     1561 gcagccgcct ggcagcacaa cagcagatgg aaggtctgct ggcaccacac cgcccaaaag
     1621 aacctgcttg gttcctggca actgtgggtg tgagccctga ccaccagggt aagggcctgg
     1681 gctctgcagt ggtgctgcct ggtgtggaag cagctgaacg tgcaggtgtg cctgctttcc
     1741 tggagacctc agctccacgc aacctgcctt tctatgaacg cctgggcttc actgtgactg
     1801 ctgatgtgga agtgccagaa ggcccacgca cttggtgcat gactcgcaaa ccaggtgcta
     1861 gccttggctc cggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     1921 ccggcccact cggctacccc tacgacgtgc ccgactacgc cagcggcatg gctaagactt
     1981 ccgaacagag ggtgaacatt gctacactgc tgacagaaaa taagaagaaa atcgtggata
     2041 aggcttccca ggatctgtgg cggagacacc cagacctgat cgcaccagga ggaattgctt
     2101 tctctcagag ggaccgcgct ctgtgcctgc gagattacgg ctggttcctg catctgatca
     2161 ccttttgtct gctggccgga gataagggcc ccatcgagtc tattgggctg atcagtattc
     2221 gagaaatgta taactcactg ggagtgcccg tccctgcaat gatggagagc attagatgcc
     2281 tgaaagaagc cagcctgtcc ctgctggacg aagaggacgc caacgagacc gcaccctact
     2341 ttgattacat tattaaggct atgtccagcc ccaagaagaa gcgcaaggtg ggcgcctaag
     2401 gctagagttc gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact
     2461 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2521 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2581 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2641 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2701 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2761 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2821 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2881 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2941 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3001 gcctccccgc ctggaattaa ttctggctaa gcacttcgtg gccgtcgatc gtttaaaggg
     3061 aggtagtgag tcgagatatc gatactagtg atccagacat gataagatac attgatgagt
     3121 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3181 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3241 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3301 tctacaaatg tggtatggct gattatgatc ctctagaggg acagcccccc cccaaagccc
     3361 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     3421 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     3481 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     3541 cagacacctg gggggatacg gggaaaagcc ataccaatgg agctcacggg gacagccccc
     3601 ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg
     3661 cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg
     3721 acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct
     3781 ctttgagcct gcagacacct ggggggatac ggggaaaaag aattcgataa aagttttgtt
     3841 actttataga agaaattttg agtttttgtt ttttttaata aataaataaa cataaataaa
     3901 ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt aatatctatt
     3961 caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca attttacaca
     4021 tgattatctt taacgtacgt cacaatatga ttatctttct agggttaatc tagctgcgtg
     4081 ttctgcagcg tgtcgagcat cttcatctgc tccatcacgc tgtaaaacac atttgcaccg
     4141 cgagtctgcc cgtcctccac gggttcaaaa acgtgaatga acgaggcgcg ctcactggcc
     4201 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4261 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4321 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4381 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4441 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4501 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4561 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4621 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4681 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4741 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4801 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4861 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4921 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4981 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5041 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5101 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5161 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5221 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5281 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5341 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5401 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5461 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5521 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5581 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5641 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5701 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5761 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5821 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5881 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6721 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6781 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6841 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6901 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6961 atgattacgc caagcgcgcc cgccgggtaa ctcacggggt atccatgtcc atttctgcgg
     7021 catccagcca ggatacccgt cctcgctgac gtaatatccc agcgccgcac cgctgtcatt
     7081 aatctgcaca ccggcacggc agttccggct gtcgccggta ttgttcgggt tgctgatgcg
     7141 cttcgggctg accatccgga actgtgtccg gaaaagccgc gacgaactgg tatcccaggt
     7201 ggcctgaacg aacagttcac cgttaaaggc gtgcatggcc acaccttccc gaatcatcat
     7261 ggtaaacgtg cgttttcgct caacgtcaat gcagcagcag tcatcctcgg caaactcttt
     7321 ccatgccgct tcaacctcgc gggaaaaggc acgggcttct tcctccccga tgcccagata
     7381 gcgccagctt gggcgatgac tgagccggaa aaaagacccg acgatatgat cctgatgcag
     7441 ctaga
//
