LOCUS       sequence-450389-        4082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty MTK PB cargo backbone with no marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4082)
  AUTHORS   Loveless BT, Carlson CK, Dentzel Helmy CA, Hu VJ, Ross SK, Demelo 
            MC, Murtaza A, Liang G, Ficht M, Singhai A, Pajoh-Casco MJ, Liu CC
  TITLE     Open-ended molecular recording of sequential cellular events into 
            DNA
  JOURNAL   bioRxiv 2021.11.05.467507
REFERENCE   2  (bases 1 to 4082)
  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: "bioRxiv 
            2021.11.05.467507"
COMMENT     Sequence Label: pTBL2367 MTK PB empty backbone with no marker
FEATURES             Location/Qualifiers
     source          1..4082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      70..141
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      157..184
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     repeat_region   780..842
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(893..911)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(951..1766)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(951..1766)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1674..1693
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1758..1777
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1991..2579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2480..2499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2691..2710
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2691..2709
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   2739..2773
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     RBS             3409..3420
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3427..4082,1..58)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaatg
       61 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      121 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
      181 tgcgtttata gctggaaatc tgctcgtcag tggtgctcac actgacgaat catgtacaga
      241 tcataccgat gactgcctgg cgactcacaa ctaagcaaga cagccggaac cagcgccggc
      301 gaacaccact gcatatatgg catatcacaa cagtccacgt ctcaagcagt tacagagatg
      361 ttacgaaccc tgaacaggtt gactattaat tgtgtctgct tgatgtggac accaggtggc
      421 gctggacatc agatttggag aggcagttgt ctagggaacc gggctctgtg ccagcgcagg
      481 aggcaggctg gctctcctat tccagggatg ctcatccagg aaggaaaggt tgcatgctgg
      541 acacactaac cttacagcgg tgggctctat ggggtaccca cgcatggtat ggatgaacac
      601 tagttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt ttttaataaa
      661 taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt gtaaatataa
      721 taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc gataaaacac
      781 atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt atctttctag
      841 ggttaatcta gtatacgcgt atgcggccgc ttaattaatc cagcttttgt tccctttagt
      901 gagggttaat tccgagcggc cgcgtgttac aaccaattaa ccaattctga ttagaaaaac
      961 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1021 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1081 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1141 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1201 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1261 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1321 aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1381 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1441 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1501 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1561 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1621 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1681 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa
     1741 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     1801 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     1861 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagtcatga
     1921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2101 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2581 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2641 tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat taatacgact
     2701 cactataggg cgaattgggg cgcgccattc tagattaacc ctagaaagat agtctgcgta
     2761 aaattgacgc atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg
     2821 ctgtgcattt aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg
     2881 cggtaagtgt cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta
     2941 tcttttacgt gacttttaag atttaactca tacgataatt atattgttat ttcatgttct
     3001 acttacgtga taacttatta tatatatatt ttcttgttat agatatcaac tagaatgcta
     3061 gcatgggccc agatctccat gggccctgaa ttcgcatcta gactgatgag acgtggtaga
     3121 gccacaaaca gccggtacaa gcaacgatct ccaggaccat ctgaatcatg cgcggatgac
     3181 acgaactcac gacggcgatc acagacatta acccacagta cagacactgc gacaacgtgg
     3241 caattcgtcg caatacaacg gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt
     3301 gtaaatctta tttaataatg tgtgcggcaa ttcacattta atttatgaat gttttcttaa
     3361 catcgcggca actcaagaaa cggcaggttc ggatcttagc tactagagaa agaggagaaa
     3421 tactagatgc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg
     3481 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact
     3541 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt tccttggccg
     3601 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg
     3661 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt
     3721 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc
     3781 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aggacggcaa tatcctgggc
     3841 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa
     3901 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg
     3961 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat
     4021 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg
     4081 gt
//
