LOCUS       sequence-444806-        5573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of MARWOLEN1 transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5573)
  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 5573)
  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: pCMV-MARWOLEN1
FEATURES             Location/Qualifiers
     source          1..5573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1761..1779)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1762..1779)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1799..1816)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1805..2029
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2075..2503
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2162..2181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2372..2393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2512..2532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2517..2846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2697..2832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2759..2778
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3012..3133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3049..3068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3103..3122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3182..3198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3182..3198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3195..3217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3206..3222
                     /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(3230..3236,3237..3254,3255..3260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3275..3296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3413..3430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3584..4172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3664..3683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4343..5134,5135..5203))
                     /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     4966..4985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5204..5308)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5383..5402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccgccaccat
      721 gcgtaaactg agcggcgaga tccagaacaa cattgtgagc ctgaccgagt caggcaacag
      781 cactcgtcag atagcggaac gtctgggcat tagccaaagc gcggtggtga gcattcagaa
      841 gcgccgcaaa ctgaccccga aacctcaagt tagcggccgt aaaaagctgc tgaaggatag
      901 cgatgcgcgc ctgatgatga gcgaaatgcg caaaaacaaa aacctgaccc ctaagggtgc
      961 atgcctggcg attaacaaaa acgtgagcga atggaccgcg cgtcgtgcct tacaggatat
     1021 cggctatatg agcatcgtga agaaaaacaa accggcgctg agcgacaaga atgtgaaagc
     1081 gcgcctgaaa tttgccaagg atcacaaaaa ctggaccatt gatgactgga agcgcgtggt
     1141 ttggagcgac gaaagcaaat tcaaccgctt ccagagcgac ggcaagcagt attgctggat
     1201 tcgccctggt gatcgtgttc agcgccatca tgtgaaacag accgtgaaac atggcggcgg
     1261 caacataatg gtgtggggct gctttacctg gtggcatatt ggtccgctgc agctggtgga
     1321 aggcattatg aagaaggaag actatctgcg cattctgcag accaacctgc cgaactattt
     1381 cgataaatgc gcgtatccgg aaaaggatat catctttcag caggatggcg atcctaaaca
     1441 caccgcgaaa attgtgaaag agtggatcgg caaacagcat tttcagctga tggagtggcc
     1501 tgcgcaaagc cctgatctga accctattga gaacctgtgg agcattgtga aacgccgtct
     1561 gggccagtat gatagcgcgc cgaagaacat gggcgatctg tgggaacgtg tggctgtgga
     1621 atggagccgt attccgcaag acattctgcg caacctggtg gaaagcatgc cgaaacgcgt
     1681 gaccgaagtg attgtgaaca agggcctgtg gaccaaatac tgatgagcgg ccgctcgagc
     1741 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     1801 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     1861 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     1921 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     1981 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2041 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2281 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2401 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2521 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2581 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2641 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2701 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2761 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     2821 tttttggagg cctaggcttt tgcaaaaagc tccccgaaat gaccgaccaa gcgacgccca
     2881 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     2941 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3001 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3121 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5341 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     5401 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     5461 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     5521 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
