LOCUS       sequence-477113-        3724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid for synthetic DNA Tape (Typewriter 5X- Tape).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3724)
  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 3724)
  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: P024-mPB-5xTAPE-BsaI Map
FEATURES             Location/Qualifiers
     source          1..3724
                     /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   309..343
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(834..850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(834..850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(847..869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    858..874
                     /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(882..888,889..906,907..912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    927..948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1065..1082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1236..1824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1316..1335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1995..2786,2787..2855))
                     /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     2618..2637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2856..2960)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2986..3441)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3123..3144)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3335..3354
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3568..3590
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3582..3599
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3583..3599
                     /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 tcagcatgga tgatggtgag
      181 cacgtgatgg tgagcacgtg atggtgagca cgtgatggtg agcacgtgat ggtgagcacg
      241 tgatggtgag cgctttaagg ccggtcctag caattggggc acttggtagt atgtgcagct
      301 cctgcaccca tgcgtcaatt ttacgcagac tatctttcta gggttaatct agctgcatca
      361 ggatcatatc gtcgggtctt ttttccggct cagtcatcgc ccaagctggc gctatctggg
      421 catcggggag gaagaagccc gtgccttttc ccgcgaggtt gaagcggcat ggaaagagtt
      481 tgccgaggat gactgctgct gcattgacgt tgagcgaaaa cgcacgttta ccatgatgat
      541 tcgggaaggt gtggccatgc acgcctttaa cggtgaactg ttcgttcagg ccacctggga
      601 taccagttcg tcgcggcttt tccggacaca gttccggatg gtcagcccga agcgcatcag
      661 caacccgaac aataccggcg acagccggaa ctgccgtgcc ggtgtgcaga ttaatgacag
      721 cggtgcggcg ctgggatatt acgtcagcga ggacgggtat cctggctgga tgccgcagaa
      781 atggacatgg ataccccgtg agttacccgg cgggcgcgct tggcgtaatc atggtcatag
      841 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
      901 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
      961 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1021 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1861 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1921 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1981 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2041 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2101 ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat
     2161 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2221 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2281 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2341 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2401 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2461 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2521 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2581 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2641 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2701 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2761 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2821 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2881 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2941 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     3001 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3061 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3121 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3181 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3241 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3301 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3361 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3421 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     3481 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3541 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     3601 agcgcgcctc gttcattcac gtttttgaac ccgtggagga cgggcagact cgcggtgcaa
     3661 atgtgtttta cagcgtgatg gagcagatga agatgctcga cacgctgcag aacacgcagc
     3721 taga
//
