LOCUS       sequence-443643-        4960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of MARISP1 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4960)
  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 4960)
  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-MARISP1
FEATURES             Location/Qualifiers
     source          1..4960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(660..689)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        1120..1619
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1155..1178)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1552..1571
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1633..1642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1639..2238
                     /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     2138..2158
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2245..2301
                     /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             join(2302..2304,2305..2307,2308..3021)
                     /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             join(2302..2304,2305..2307,2308..3018)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2347..2368)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2608..2627)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2955..2976
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3043..3060)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3049..3273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3664..3680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3664..3680)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3692..3709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3761..3792
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3793..3895
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3896..3964,3965..4756)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4114..4133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4757..4948
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaataggc gtatcacgag gccaataggc gtatcacgag gtctctctct actagttata
      781 cactgtcgct cataaagttg gaatattttt ttttttaatc aaaccccgta catttatcaa
      841 tttttacgct tcagtaatca ttatatgcca atttttttaa ttgttcaatg tgtctccact
      901 accaggtgta atgaatagtt gcataataat gcattccatg agccttttgt ttgaattctt
      961 gtgtttgaag ctacaagagg catcgcgctt cgatattttg tccattttgc tctcataggc
     1021 ttatgtgtaa caagttcact tttttcagag ttggctgtat cattttcaac aaaatttggt
     1081 acagaggttc tctaagccac cggtgctagc aattctaccg ggtaggggag gcgcttttcc
     1141 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     1201 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     1261 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     1321 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     1381 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     1441 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     1501 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     1561 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     1621 cctgcagcca acgccaccat ggggaccgag tacaagccca cggtgcgcct cgccacccgc
     1681 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     1741 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     1801 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     1861 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     1921 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     1981 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     2041 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     2101 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     2161 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     2221 acccgcaagc ccggtgccat cgatgccact aacttctccc tgttgaaaca agcaggggat
     2281 gtcgaagaga atcccgggcc aatggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaagta attaattaag ctcgctgatc agcctcgact gtgccttcta
     3061 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3121 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3181 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3241 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcgcgccct cgagtgactg
     3301 attgacattt gaaagagaaa acagaacaca tttcatgcaa aaaaatcata taaatatttg
     3361 ttatctgtta tgcagaccaa tttttaacat tgtaacagaa tccttagttt tgctatgtcc
     3421 ataccccttt ttttgtttct gccttgatac accctttaca aagggtaatt gtagtttaat
     3481 ttcactgaaa aatgacctga aggaatcgat caataaaaca ttatggttgt ttacaccata
     3541 gatatgataa ataacgcgta atgtcttatt tattgtacgt taggttgtaa aaaaaaaaaa
     3601 tattccaact ttatgagcga cagtgtatag ggcccagaga cggagtcact gccaaccgag
     3661 acggtcatag ctgtttcctg tgtgccgctt cctcgctcac tgactcgctg cgctcggtcg
     3721 ttactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg
     3781 aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     3841 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag
     3901 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3961 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4021 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagtttac gccccgaaga
     4081 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4141 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4201 gtactcacca gtcacagaaa agcatctcac ggatggcatg acagtaagag aattatgcag
     4261 tgctgccata accatgagtg ataacactgc ggccaactta cttctggcaa cgatcggagg
     4321 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4381 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4441 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4501 gcaacaatta atagactgga tggaggcgga taaagttgca ggatcacttc tgcgctcggc
     4561 cctcccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg
     4621 tatcattgca gaactggggc cagatggtaa gccctcccgc atcgtagtta tctacacgac
     4681 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4741 gattaagcat tggtaagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     4801 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     4861 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     4921 tttcggtgat gacggtgaaa acctctgatg agggcccaaa
//
