LOCUS       sequence-444318-        5963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Mariner-2_PM 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  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 5963)
  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-Mariner-2_PM
FEATURES             Location/Qualifiers
     source          1..5963
                     /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"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2151..2169)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2152..2169)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2189..2206)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2195..2419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2465..2893
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2552..2571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2762..2783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2902..2922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2907..3236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3087..3222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3149..3168
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3402..3523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3439..3458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3493..3512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3572..3588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3572..3588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3585..3607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3596..3612
                     /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(3620..3626,3627..3644,3645..3650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3665..3686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3803..3820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3974..4562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4054..4073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4733..5524,5525..5593))
                     /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     5356..5375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5594..5698)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5773..5792)
                     /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 ggcccctaaa aaagtgagcg gcagcgataa accgaaacgc aaagtgctgc gcaccaccat
      781 tgagctgcgc aaagagatta tcgaaaaggc ggaacgtggt gtgcgtgtga ccgatatcag
      841 caaggaatat ggcatgccta aaagcaccat ttgcaccatc ctgaagaaca aagaggccat
      901 taaggaagcg aacgtggcga aaggtgttag cagcctgagc aagcaacgca ccaaagcgat
      961 cgaggaagtg gaaaaactgc tgctgatctg gattaacgaa aaacagctgg cgggcgatag
     1021 cgtttcagag gcgatgatct gcgaaaaggc gaagcagctg catagcgacc tgctgaaaaa
     1081 taccccgggc acctcagatg atgcgggccc gagcgacatt tttaaagcga gccgcggctg
     1141 gttcgataaa ttcaaaaagc gcagcggcat tcatagcgtg gtgcgccatg gtgaagcggc
     1201 gagcgcgaat aaagaagccg ccaagaactt cgtgggcgag tttaaagagt ttgtggatgc
     1261 ggaaggcttt atccctcagc aggtgtttaa ctgcgatgaa accggcctgt tttggaagaa
     1321 aatgccgcgc cgcacctata ttacccagga agaaaaagcg ctgccgggcc ataaacctat
     1381 gaaggaccgc ctgaccctgc tgttatgtgg caacgcgagc ggtgacttca aagtgaaacc
     1441 gctgctggtg taccatagcg aaaaccctcg cgtgtttaaa aagaacaacg tgatgaagaa
     1501 caagctgcat gtgatgtggc gcgccaatag caaagcatgg gtgacccgcc agttttttat
     1561 tgaatgggtg catgaggtgt ttggcccgag cgtgaagaaa tacctgcagg aaaagcagct
     1621 gcctctgaag gcgctgctgt taatggataa cgcccctgca catcctcctg gcttagaaga
     1681 atcatgccgc ggcagctttc tgattaccgt gaagttcctg cctccgaaca ccacccctct
     1741 gattcagcct atggaccagc aggtgatcag caactttaaa aagctgtaca ccaaagcgct
     1801 gttccagcgc tgtttcgaag tgaccagcga caccgaatta accctgcgcg agttctggaa
     1861 gaaccacttc aacattctgc attgcctgag cctgatcgat aaggcgtggg gcgaagtttc
     1921 aagccgcacc atgaatagcg cgtggcgtaa actgtggccg gattgtgttg cggagcgcga
     1981 ttttgaaggc tttgaaccgg aacctgagac tgaaccggtg gtggaaaaca ttgtgagcct
     2041 gggcaagtca atgggcctgg aagtggatga tgaaatgctg cgcagctggt ggcgtaccac
     2101 cgcccagaac taatgagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     2161 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2221 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2281 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2341 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2401 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2461 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2521 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2581 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2641 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2701 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2761 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2821 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2881 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3121 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3181 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3241 tccccgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3301 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3361 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3421 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3481 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3541 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3601 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3661 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3721 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4381 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4621 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4681 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4741 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4801 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4861 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4921 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4981 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5041 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5101 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5161 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5221 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5281 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5341 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5401 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5461 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5521 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5581 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5701 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     5761 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     5821 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     5881 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     5941 tgtacgggcc agatatacgc gtt
//
