LOCUS       sequence-434144-        4668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac, constitutive expression destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4668)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 4668)
  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"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pRDA_575
FEATURES             Location/Qualifiers
     source          1..4668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      656..665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             662..1339
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     polyA_signal    1382..1503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1419..1438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1473..1492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1775..1809
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1874..1890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1874..1890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1887..1909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1898..1914
                     /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)."
     promoter        complement(join(1922..1928,1929..1946,1947..1952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1967..1988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2105..2122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2276..2864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2356..2375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3035..3826,3827..3895))
                     /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     3658..3677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3896..4000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4068..4086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4124..4146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4246..4265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4459..4481
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4474..4490
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   4549..4611
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 tattatattt acacttacat actaataata aattcaacaa acaatttatt tatgtttatt
       61 tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttatga
      121 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
      181 agcagcgagc cgcccccggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
      241 cagcgtgcgg ggacagcccg cgcacgggga aggtggcacg ggatcgcttt cctctgaacg
      301 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa ggcctccagg
      361 tacctacacg cgtcttccat gcgggcagag cgcacatcgc ccacagtccc cgagaagttg
      421 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
      481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaggt ctatataagc
      541 agagctcgtt tagtgaaccg tcagtccgcc tggagacctc gagccgagtg gtcgtgcctc
      601 catagaaaga ataaactagt attcttctgg tccccacaga ctcagagaga accccgccac
      661 catggatagc actgagaacg tcatcaagcc cttcatgcgc ttcaaggtgc acatggaggg
      721 ctccgtgaac ggccacgagt tcgagatcga gggcgtgggc gagggcaagc cctacgaggg
      781 cacccagacc gccaagctgc aagtgaccaa gggcggcccc ctgcccttcg cctgggacat
      841 cctgtccccc cagttcttct acggctccaa ggcgtacatc aagcaccccg ccgacatccc
      901 cgactacctc aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      961 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggca ccctcatcta
     1021 ccacgtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaaaaagac
     1081 tctgggctgg gagccctcca ctgagcgcaa ctacccccgc gacggcgtgc tgaagggcga
     1141 gaaccacatg gcgctgaagc tgaagggcgg cggccactac ctgtgtgagt tcaagtccat
     1201 ctacatggcc aagaagcccg tgaagctgcc cggctaccac tacgtggact acaagctcga
     1261 catcacctcc cacaacgagg actacaccgt ggtggagcag tacgagcgcg ccgaggcccg
     1321 ccaccacctg ttccagtaaa tgcatgaaga cgcattcgaa ttccatgcct ttgtatcagt
     1381 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1501 ttaatctata acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata
     1561 acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca
     1621 ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac
     1681 gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg
     1741 ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc
     1801 tttctagggt taatctatcg gatcccgggc ccgtcgactg cagaggcctg catgcaagct
     1861 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1921 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1981 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2041 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2101 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2161 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2221 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2281 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2341 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2401 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2461 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2521 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2581 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2641 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2701 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2761 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2821 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2881 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2941 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3001 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3061 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3121 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3181 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3241 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3301 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3361 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3421 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3481 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3541 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3601 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3661 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3721 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3781 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3841 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3901 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3961 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4021 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4081 cgaggccctt tcgtcttgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4141 tcccggaaac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4201 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4261 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4321 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4381 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4441 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     4501 tcggtacctc gcgaatgcat ctagatatcg acggccagtc ccgattaacc ctagaaagat
     4561 aatcatattg tgacgtacgt taaagataat catgtgtaaa attgacgcat gtgttttatc
     4621 ggtctgtata tcgaggttta tttattaatt tgaatagata ttaagttt
//
