LOCUS       sequence-256380-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression of iRFP702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 7944)
  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-iRFP702
FEATURES             Location/Qualifiers
     source          1..7944
                     /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"
     misc_feature    2921..3509
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2974..2994)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3392..3403)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3589..3723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3613..3632)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3667..3686
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4501..4563
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4693..4710)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4693..4709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4702..4724)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4851..5306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4938..4957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5148..5169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5332..5436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5437..5505,5506..6297)
                     /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(5655..5674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6468..7056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6957..6976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7210..7227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7344..7365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7380..7385,7386..7403,7404..7410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7418..7434
                     /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     7423..7445
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7442..7458
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7442..7458
                     /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 gccagaaagg
     1981 tggatctgac aagctgtgac agagaaccca tccacattcc tggcagcatc caaccctgtg
     2041 gctgcctcct ggcttgtgat gcacaagcag tcaggattac aaggattaca gaaaatgctg
     2101 gagccttctt tggaagggaa acccctaggg tgggagagct gttggccgat tactttggcg
     2161 aaacagaggc ccatgccttg agaaacgcac tggctcagag ctctgatcct aaaaggcctg
     2221 ctctgatctt tggctggaga gatggcctca ctggcagaac cttcgatatt agcctccata
     2281 ggcacgatgg aacctccatt attgaatttg agcccgctgc cgctgagcag gctgataacc
     2341 ctctgagact cacaagacaa atcattgcca ggaccaagga actcaagtcc ctggaagaaa
     2401 tggcagccag ggtgccaaga tatctccagg ctatgctggg ctaccacaga gtcatgctgt
     2461 ataggtttgc tgatgatgga agcgggaagg tcattgggga ggcaaagagg tctgacctcg
     2521 aaagcttctt gggacaacac ttccctgcca gcctggtccc ccagcaggcc aggctgctct
     2581 atctcaagaa tgccatcagg gtggtgtctg atagcagagg gatcagctcc agaatcgtcc
     2641 ctgagcatga cgcctctggc gctgccctgg atttgagctt tgctcacctc aggagcatct
     2701 ctcctataca tttggaattt ctgaggaaca tgggggtgtc tgcctccatg agcctgagca
     2761 tcattatcga tggcaccctc tggggcctca ttatctgcca ccactatgaa cctagagctg
     2821 tccctatggc acagagagtg gctgctgaga tgttcgctga ttttctcagc ttgcacttta
     2881 ctgctgctca tcaccaaaga taagagttcg cgtctggaac aatcaacctc tggattacaa
     2941 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3001 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3061 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3121 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3181 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3241 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3301 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3361 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3421 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3481 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctggctaag cacttcgtgg
     3541 ccgtcgatcg tttaaaggga ggtagtgagt cgagatatcg atactagtga tccagacatg
     3601 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3661 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3721 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3781 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagaggga
     3841 cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca
     3901 gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc
     3961 gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc
     4021 tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaagcca taccaatgga
     4081 gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct
     4141 agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc
     4201 gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct
     4261 ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaaga
     4321 attcgataaa agttttgtta ctttatagaa gaaattttga gtttttgttt tttttaataa
     4381 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata
     4441 ataaaactta atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca
     4501 catgcgtcaa ttttacacat gattatcttt aacgtacgtc acaatatgat tatctttcta
     4561 gggttaatct agctgcgtgt tctgcagcgt gtcgagcatc ttcatctgct ccatcacgct
     4621 gtaaaacaca tttgcaccgc gagtctgccc gtcctccacg ggttcaaaaa cgtgaatgaa
     4681 cgaggcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4741 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4801 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     4861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4981 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5041 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5161 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5221 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5281 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     5341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5401 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5461 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5521 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5581 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5641 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5701 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5761 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5821 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5881 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5941 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6001 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6061 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6121 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6181 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6241 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6301 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6601 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7081 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7141 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7201 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7261 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7321 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7381 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7441 acaggaaaca gctatgacca tgattacgcc aagcgcgccc gccgggtaac tcacggggta
     7501 tccatgtcca tttctgcggc atccagccag gatacccgtc ctcgctgacg taatatccca
     7561 gcgccgcacc gctgtcatta atctgcacac cggcacggca gttccggctg tcgccggtat
     7621 tgttcgggtt gctgatgcgc ttcgggctga ccatccggaa ctgtgtccgg aaaagccgcg
     7681 acgaactggt atcccaggtg gcctgaacga acagttcacc gttaaaggcg tgcatggcca
     7741 caccttcccg aatcatcatg gtaaacgtgc gttttcgctc aacgtcaatg cagcagcagt
     7801 catcctcggc aaactctttc catgccgctt caacctcgcg ggaaaaggca cgggcttctt
     7861 cctccccgat gcccagatag cgccagcttg ggcgatgact gagccggaaa aaagacccga
     7921 cgatatgatc ctgatgcagc taga
//
