LOCUS       sequence-340874-        6279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes 5xTAPE-1 construct that can be integrated using piggyBAC 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6279)
  AUTHORS   Choi J, Chen W, Minkina A, Chardon FM, Suiter CC, Regalado SG, 
            Domcke S, Hamazaki N, Lee C, Martin B, Daza RM, Shendure J
  TITLE     A time-resolved, multi-symbol molecular recorder via sequential 
            genome editing.
  JOURNAL   Nature. 2022 Jul 6. pii: 10.1038/s41586-022-04922-8. doi: 
            10.1038/s41586-022-04922-8.
  PUBMED    35794474
REFERENCE   2  (bases 1 to 6279)
  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: "Nature. 
            2022 Jul 6. pii: 10.1038/s41586-022-04922-8. doi: 
            10.1038/s41586-022-04922-8."
COMMENT     Sequence Label: piggyBAC-5xTAPE-1-BlastR
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1361..1609
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1361..1381
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1532..1551
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        1803..2014
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2027..2295
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2323..2332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2329..2727
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(2787..2807)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2849..2970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2886..2905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2940..2959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3733..3767
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4258..4274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4258..4274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4271..4293)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4282..4298
                     /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(4306..4312,4313..4330,4331..4336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4489..4506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4660..5248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4740..4759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5419..6210,6211..6279))
                     /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     6042..6061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccaaga gagggcctat ttcccatgat
     1381 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac
     1441 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     1501 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     1561 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccn nnnnnnngga
     1621 tgatggtgag cacgtgatgg tgagcacgtg atggtgagca cgtgatggtg agcacgtgat
     1681 ggtgagcacg tgatggtgag cgctttaagg ccggtcctag caattttttt aagcttggcg
     1741 taactagatc ttgagacaaa cgcggccgcg aaggatctgc gatcgctccg gtgcccgtca
     1801 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     1861 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     1921 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     1981 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     2041 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     2101 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     2161 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     2221 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     2281 gcgccgttac agatccaagc tgtgaccggc gcctaccgta cggccaccat ggccaagcct
     2341 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     2401 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     2461 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     2521 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     2581 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     2641 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     2701 ccctctggtt atgtgtggga gggctaaacg cgtaatcaac ctctggatta caaaatttgt
     2761 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2821 ttaatgcctt tgtatcatgc gttaactaaa cttgtttatt gcagcttata atggttacaa
     2881 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2941 tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca
     3001 attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc cctgtttaat
     3061 atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt
     3121 gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta
     3181 gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt tccccgtatc
     3241 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt
     3301 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag
     3361 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt
     3421 aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat
     3481 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     3541 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     3601 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     3661 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     3721 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc
     3781 atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc
     3841 tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag
     3901 agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga
     3961 tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct
     4021 gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca
     4081 tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg
     4141 acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc
     4201 agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc
     4261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5101 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6241 aaagggaata agggcgacac ggaaatgttg aatactcat
//
