LOCUS       sequence-443033-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Mariner2_AG 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  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 5570)
  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-Mariner2_AG
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        448..651
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     602..622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      780..789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1825..1843)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1826..1843)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1863..1880)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1869..2093
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2139..2567
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2226..2245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2436..2457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2576..2596)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2581..2910
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2761..2896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2823..2842
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3076..3197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3113..3132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3167..3186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3246..3262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3246..3262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3259..3281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3270..3286
                     /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(3294..3300,3301..3318,3319..3324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3339..3360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3477..3494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3648..4236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3728..3747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4407..5198,5199..5267))
                     /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     5030..5049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5268..5372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5447..5466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
       61 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      661 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      721 caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg gaattcgccg
      781 ccaccatggg tcgttcagcg catagcaccc aacaacagcg cctggacatt aaacgcttaa
      841 gcgccgccgg ttatagccaa cgcaaaattg cggaaatcct gggccgcagc aagacctttg
      901 tgtataacgc cctgcatagc accggcacca aaattcctac tggccgccct cgtaaaacta
      961 gcgcgcgtga tgatgcacgt atgacccgtc tgtgtaaagc cgacccgttt aaaagcgcgc
     1021 gcgccattcg tgatgaactg cagctgagcg ttagcgatcg caccgttcaa cgtcgcctgt
     1081 tcgagaataa tctggtgggc cgcaatcctc gtaaagtgcc tctgctgcgt cgttgtcatg
     1141 tgcaggcacg cttacaattt gcgcgcgagc actatgattg ggcgggcgag aacttaaaca
     1201 aatggcgcaa cgtgctgtgg agcgatgaga gcaaagtgaa cctggtgggc agcgatggca
     1261 aacgttttgt gcgccgccct aagaataccg cgtatcgccc tcagtatacc ctgaaaaccg
     1321 tgaaacatgg cggcggcaac attatggtgt gggcgtgctt ctcatggtat ggcgtgggcc
     1381 ctatcttctg gatcaaagac atcatggatc agcatcgcta cctgaacatt attcagaccg
     1441 tgatgctgcc tcacgccgaa tgggaaatgc ctctgaagtg gcagttcatg catgataacg
     1501 acccgaagca taccgccaaa gcggtgaaga aatggttcgt ggaccagaag attgatgtga
     1561 tgaactggcc tgcgcagagc cctgatctga acccgatcga aaacctgtgg aagattgtga
     1621 aggccaagct gcctcctgca ggttcacgca ccaaagagaa actgtggaag catattgaaa
     1681 acgcgtggta cagcatacct ccgagcacct gtaaaagcct ggtggaaagc atgccgaaac
     1741 gtatgcgcgc ggtgattcgc aataacggcc atgcgaccaa atattgatga gcggccgctc
     1801 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     1861 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1921 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1981 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2041 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2101 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2161 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2221 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2281 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2341 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2401 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2461 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2581 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2641 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2701 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2761 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2821 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2881 ggcttttttg gaggcctagg cttttgcaaa aagctccccg aaatgaccga ccaagcgacg
     2941 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3001 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3061 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3181 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5401 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     5461 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     5521 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
//
