LOCUS       sequence-477112-        3727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid for synthetic DNA Tape (HEK3 locus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3727)
  AUTHORS   Chen W, Choi J, Li X, Nathans JF, Martin B, Yang W, Hamazaki N, Qiu 
            C, Lalanne JB, Regalado S, Kim H, Agarwal V, Nichols E, Leith A, Lee
            C, Shendure J
  TITLE     Symbolic recording of signalling and cis-regulatory element activity
            to DNA.
  JOURNAL   Nature. 2024 Aug;632(8027):1073-1081. doi: 
            10.1038/s41586-024-07706-4. Epub 2024 Jul 17.
  PUBMED    39020177
REFERENCE   2  (bases 1 to 3727)
  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"; doi: 
            "10.1038/s41586-024-07706-4"; journalName: "Nature"; date: 
            "2024-08"; volume: "632"; issue: "8027"; pages: "1073-1081"
COMMENT     Sequence Label: P023-mPB-synHEK3-BsaI
FEATURES             Location/Qualifiers
     source          1..3727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..67
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   312..346
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(837..853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(837..853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(850..872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    861..877
                     /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(885..891,892..909,910..915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    930..951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1068..1085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1239..1827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1319..1338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1998..2789,2790..2858))
                     /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     2621..2640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2859..2963)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2989..3444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3126..3147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3338..3357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3571..3593
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3585..3602
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3586..3602
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg tgtaaaattg
       61 acgcatgtgt tttatcggtc tgttaatacg actcactata ggatatcggc tatatcttgt
      121 ggaaaggacg aaacacccga gaccctgcca atgataggtc tcagcatctc cagccctggc
      181 ctgggtcaat ccttggggcc cagactgagc acgtgatggc agaggaaagg aagccctgct
      241 tcctccagag ggcgtcgcag gacagctttt cctagacagg ggcacttggt agtatgtgca
      301 gctcctgcac ccatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca
      361 tcaggatcat atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct
      421 gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga
      481 gtttgccgag gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat
      541 gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg
      601 ggataccagt tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat
      661 cagcaacccg aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga
      721 cagcggtgcg gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca
      781 gaaatggaca tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca
      841 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      901 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      961 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1021 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1081 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1141 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1201 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1261 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1321 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1381 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1441 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1501 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1561 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1621 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1681 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1741 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1801 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1861 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1921 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1981 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2041 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2101 ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca
     2161 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2221 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2281 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2341 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2401 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2461 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2521 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2581 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2641 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2701 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2761 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2821 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2881 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2941 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     3001 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3061 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3421 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     3481 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3541 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3601 gtgagcgcgc ctcgttcatt cacgtttttg aacccgtgga ggacgggcag actcgcggtg
     3661 caaatgtgtt ttacagcgtg atggagcaga tgaagatgct cgacacgctg cagaacacgc
     3721 agctaga
//
