LOCUS       sequence-444746-        4714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-1_PM Transposon without spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4714)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 4714)
  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: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pMV-Tc1-1_PM w/o spacer
FEATURES             Location/Qualifiers
     source          1..4714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      279..288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(302..801)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(350..369)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     743..766
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(1072..1089)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1072..1088)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1098..1116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1184..1288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1289..1357,1358..2149)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1507..1526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2320..2908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2809..2828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3062..3079
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3091..3107
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3091..3107
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3362..3586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3575..3592
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3614..4327,4328..4330,4331..4333))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3617..4327,4328..4330,4331..4333))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3659..3680)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4008..4027
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4267..4288
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4334..4390)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(4397..4714,1..282))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4477..4497)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc
       61 caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag
      121 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccgggtcga cggtgtggcg
      181 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggcccggg ggacgtcgtc
      241 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcccc atggtggcgt tggctgcagg
      301 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg
      361 tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc
      421 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa
      481 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca
      541 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct
      601 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac
      661 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc
      721 cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca gactgccttg
      781 ggaaaagcgc ctcccctacc cggtagaatt gctagcaggg ggtcaaatac ttatttccct
      841 cattaaaatg caaatcaatt tataacattt ttgacatgcg tttttctgga tttttgttgt
      901 tgttattctg tctctcactg ttcaaataaa cctaccatta aaattataga ctgatcattt
      961 ctttgtcagt gggcaaacgt acaaaatcag caggggatca aatacttttt tccctcactg
     1021 taaagcttag agacgcaaga cactgcggat cgagacgcat tgcgtggttc gactggccgt
     1081 cgttttacaa cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     1141 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1621 tacggatggc atgacagtaa gagaattatg tagtgctgcc ataaccatga gtgataacac
     1681 ggcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgcg
     1741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1801 accaaacgac gagcgtgata ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact
     1861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1981 taaatctgga gccggtgagc gtgggtcccg cggtattatt gcagcactgg ggccagatgg
     2041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2101 aaagagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2941 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtggg
     3001 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3061 cagcgagtca gtgagcgagg aagcggcaca caggaaacag ctatgaccgt ctcggttggc
     3121 agtgactccg tctctgggcc ctacagtgag ggaaaaaagt atttgatccc ctgctgattt
     3181 tgtacgtttg cccactgaca aagaaatgat cagtctataa ttttaatggt aggtttattt
     3241 gaacagtgag agacagaata acaacaacaa aatccagaaa aacgcatgtc aaaaatgtta
     3301 taaattgatt tgcattttaa tgagggaaat aagtatttga cccctcggcg cgcctcagaa
     3361 gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg
     3421 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     3481 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     3541 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
     3601 cgagcttaat taattacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3661 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     3721 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3781 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     3841 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     3901 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3961 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     4021 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     4081 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     4141 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     4201 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     4261 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     4321 ccttgctcac cattggcccg ggattctctt cgacatcccc tgcttgtttc aacagggaga
     4381 agttagtggc atcgatggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     4441 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     4501 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     4561 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
     4621 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     4681 cggccagccg ggaaccgctc aactcggcca tgcg
//
