LOCUS       sequence-267650-        7945 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 7945)
  TITLE     Madison Lab plasmids
REFERENCE   2  (bases 1 to 7945)
  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..7945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   570..604
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(873..889)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(873..889)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2440..2493
                     /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             2500..2526
                     /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             2500..2526
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3486..4074
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3539..3559)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3957..3968)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4154..4288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4232..4251
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5067..5129
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5259..5276)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5259..5275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5290)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5417..5872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5504..5523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5714..5735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5898..6002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6003..6071,6072..6863)
                     /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(6221..6240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7034..7622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7523..7542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7776..7793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7910..7931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcgcgcc cgccgggtaa ctcacggggt
      121 atccatgtcc atttctgcgg catccagcca ggatacccgt cctcgctgac gtaatatccc
      181 agcgccgcac cgctgtcatt aatctgcaca ccggcacggc agttccggct gtcgccggta
      241 ttgttcgggt tgctgatgcg cttcgggctg accatccgga actgtgtccg gaaaagccgc
      301 gacgaactgg tatcccaggt ggcctgaacg aacagttcac cgttaaaggc gtgcatggcc
      361 acaccttccc gaatcatcat ggtaaacgtg cgttttcgct caacgtcaat gcagcagcag
      421 tcatcctcgg caaactcttt ccatgccgct tcaacctcgc gggaaaaggc acgggcttct
      481 tcctccccga tgcccagata gcgccagctt gggcgatgac tgagccggaa aaaagacccg
      541 acgatatgat cctgatgcag ctagattaac cctagaaaga tagtctgcgt aaaattgacg
      601 catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt
      661 taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg
      721 tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg
      781 tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg
      841 ataacttatt atatatatat tttcttgtta tagatccact agttctagag ggacagcccc
      901 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
      961 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
     1021 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
     1081 tctttgagcc tgcagacacc tggggggata cggggaaaag gcctccaagg cctgcaggag
     1141 tcaatgggaa aaacccattg gagccaagta cactgactca atagggactt tccattgggt
     1201 tttgcccagt acataaggtc aatagggggt gagtcaacag gaaagtccca ttggagccaa
     1261 gtacattgag tcaataggga ctttccaatg ggttttgccc agtacataag gtcaatggga
     1321 ggtaagccaa tgggtttttc ccattactga catgtatact gagtcattag ggactttcca
     1381 atgggttttg cccagtacat aaggtcaata ggggtgaatc aacaggaaag tcccattgga
     1441 gccaagtaca ctgagtcaat agggactttc cattgggttt tgcccagtac aaaaggtcaa
     1501 tagggggtga gtcaatgggt ttttcccatt attggcacat acataaggtc aataggggtg
     1561 actagtggag aagagcatgc ttgagggctg agtgcccctc agtgggcaga gagcacatgg
     1621 cccacagtcc ctgagaagtt ggggggaggg gtgggcaatt gaactggtgc ctagagaagg
     1681 tggggcttgg gtaaactggg aaagtgatgt ggtgtactgg ctccaccttt ttccccaggg
     1741 tgggggagaa ccatatataa gtgcagtagt ctctgtgaac attcggttta gtgaacctgc
     1801 aggtcggccg ccacgaccgg tgccaccatg actgaataca aaccaactgt tcgcctggca
     1861 actcgtgatg atgttccacg tgcagttcgc accctggctg ctgcatttgc tgactaccct
     1921 gcaacccgtc acactgtgga cccagaccgc cacattgaac gtgtgactga actgcaggag
     1981 ctgttcctga cccgtgtggg cctggacatt ggcaaagtgt gggtggcaga tgatggtgct
     2041 gctgtggcag tgtggaccac ccctgaatct gttgaagctg gtgcagtgtt tgctgagatt
     2101 ggcccacgca tggcagaact gtctggcagc cgcctggcag cacaacagca gatggaaggt
     2161 ctgctggcac cacaccgccc aaaagaacct gcttggttcc tggcaactgt gggtgtgagc
     2221 cctgaccacc agggtaaggg cctgggctct gcagtggtgc tgcctggtgt ggaagcagct
     2281 gaacgtgcag gtgtgcctgc tttcctggag acctcagctc cacgcaacct gcctttctat
     2341 gaacgcctgg gcttcactgt gactgctgat gtggaagtgc cagaaggccc acgcacttgg
     2401 tgcatgactc gcaaaccagg tgctagcctt ggctccggcg agggcagggg aagtcttcta
     2461 acatgcgggg acgtggagga aaatcccggc ccactcggct acccctacga cgtgcccgac
     2521 tacgccagcg gcatggccag aaaggtggat ctgacaagct gtgacagaga acccatccac
     2581 attcctggca gcatccaacc ctgtggctgc ctcctggctt gtgatgcaca agcagtcagg
     2641 attacaagga ttacagaaaa tgctggagcc ttctttggaa gggaaacccc tagggtggga
     2701 gagctgttgg ccgattactt tggcgaaaca gaggcccatg ccttgagaaa cgcactggct
     2761 cagagctctg atcctaaaag gcctgctctg atctttggct ggagagatgg cctcactggc
     2821 agaaccttcg atattagcct ccataggcac gatggaacct ccattattga atttgagccc
     2881 gctgccgctg agcaggctga taaccctctg agactcacaa gacaaatcat tgccaggacc
     2941 aaggaactca agtccctgga agaaatggca gccagggtgc caagatatct ccaggctatg
     3001 ctgggctacc acagagtcat gctgtatagg tttgctgatg atggaagcgg gaaggtcatt
     3061 ggggaggcaa agaggtctga cctcgaaagc ttcttgggac aacacttccc tgccagcctg
     3121 gtcccccagc aggccaggct gctctatctc aagaatgcca tcagggtggt gtctgatagc
     3181 agagggatca gctccagaat cgtccctgag catgacgcct ctggcgctgc cctggatttg
     3241 agctttgctc acctcaggag catctctcct atacatttgg aatttctgag gaacatgggg
     3301 gtgtctgcct ccatgagcct gagcatcatt atcgatggca ccctctgggg cctcattatc
     3361 tgccaccact atgaacctag agctgtccct atggcacaga gagtggctgc tgagatgttc
     3421 gctgattttc tcagcttgca ctttactgct gctcatcacc aaagataaga gttcgcgtct
     3481 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3541 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3601 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3661 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3721 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3781 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3841 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3901 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3961 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4021 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     4081 ttaattctgg ctaagcactt cgtggccgtc gatcgtttaa agggaggtag tgagtcgaga
     4141 tatcgatact agtgatccag acatgataag atacattgat gagtttggac aaaccacaac
     4201 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4261 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4321 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     4381 ggctgattat gatcctctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     4441 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     4501 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     4561 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     4621 tacggggaaa aagccatacc aatggagctc acggggacag ccccccccca aagcccccag
     4681 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     4741 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     4801 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     4861 cacctggggg gatacgggga aaaagaattc gataaaagtt ttgttacttt atagaagaaa
     4921 ttttgagttt ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt
     4981 attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa
     5041 acctcgatat acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg
     5101 tacgtcacaa tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg
     5161 agcatcttca tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc
     5221 tccacgggtt caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt
     5281 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5341 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5401 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5461 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5521 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5581 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5641 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5701 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5761 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5821 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     5881 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5941 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6001 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6061 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6121 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6181 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6241 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6301 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6361 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6421 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6481 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6541 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6601 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6661 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6721 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6781 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6841 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6901 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6961 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7021 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7081 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7141 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7201 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7261 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7321 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7381 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7441 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7501 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7561 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7621 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7681 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7741 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7801 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7861 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7921 agctcactca ttaggcaccc caggc
//
