LOCUS       sequence-443179-        6047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Mariner-2_Ami 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  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 6047)
  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_AMi
FEATURES             Location/Qualifiers
     source          1..6047
                     /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(2235..2253)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2236..2253)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2273..2290)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2279..2503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2549..2977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2636..2655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2846..2867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2986..3006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2991..3320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3171..3306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3233..3252
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3486..3607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3577..3596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3656..3672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3669..3691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3680..3696
                     /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(3704..3710,3711..3728,3729..3734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3749..3770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4058..4646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4817..5608,5609..5677))
                     /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     5440..5459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5678..5782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5857..5876)
                     /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 ggaacgcctg ctgatcctgt ggattcagga tcagacccag cgctgcgtgc ctttaagctt
      781 agccaccatt caggccaaag cgctgagcct gtttgatacc gtgaagcagc aacgtggcga
      841 gggcagcaaa gatgagaaat ttagcgccag caaaggctgg tttatgcgct ttaagggccg
      901 ctatgcgctg cataacatcc gtgtgcaagg tgaagcagca agcgccgatg aaatagcggc
      961 ccaggatttc cctaaaaaac tggcgaagat catcgaagaa ggcggctatt gtgcgcgtca
     1021 ggtgttcaac gtggatgaaa ccggcctgtt ctggaaaaag atgccgagcc gcacctatat
     1081 tgcgcagcag gaaaagacca tgcctggctt taaagcggcg aaggaccgct taaccttact
     1141 gctgggcagc aacgcagcgg gtgattttaa gctgaagcct ctgctggtgt accacagcga
     1201 aaatccgcgt gcgctgaaag gttatgtgaa gggcagcctg ccggtgattt ggaaaagcaa
     1261 ccgcaaggcg tgggttactg gcgcactgtt tgaggaatgg ttcaccgatt actttgtgcc
     1321 ggccgtgcag aattattgcg aacgcaacaa cctggcgcat aaagcgctgc tgatcctgga
     1381 taacgcgcct ggtcatcctg cgcatttagc cgatctggat ccgaacatca aggtggtgtt
     1441 tctgccgcct aacaccacca gcttactgca gcctatggac cagggcgtga ttagcagctt
     1501 caaagcgtat tatctgcgcc gtacctttgc acgcgccatt agcatcaccg atcaggaagg
     1561 tggtccgacc ctgaaggagt tctggaaagg cttcaacatt ctggatgcgg tgaaaaacat
     1621 tggcgaaagc tggaaggagg tgaaagaaag caacctgaac ggcgtgtgga aaaaactgtg
     1681 ccctgagttt gtgagcgatt ttcagggctt tgaagatcag gtggacgaag ttaccgcgga
     1741 cgtggttaaa atggcgggcc agctgcaatt agaagtggaa accgaagatg tggaggagct
     1801 gttagcgagc catagccagg agctgaccaa cgaagagctg gtgcgcttag aagagcaacg
     1861 caaggcggaa aaccaggcgc agattcagga aagccctgcg caaggtacca tgaccaccaa
     1921 gcagatgagc gaggccttca aacatctgga agcggcgatg gccattttcg aaaagatgga
     1981 tcctaacctg gagcgcagca gcaagaccaa caacctgatt agcaacgcgt acgcgtgcta
     2041 tcgcgagatt tacaacgaaa agaagcgcgc cgcgaaacaa actagcctgg atcagtttct
     2101 gaaaaaaccg agcagcggcc ctaaaagccc tcagaaaagc cctcagcgca gcccttttca
     2161 tgatcctggt gagggcgttc ctagccctag cgattaatga gcggccgctc gagcatgcat
     2221 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     2281 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2341 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2401 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2461 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2521 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2941 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa aagctccccg aaatgaccga ccaagcgacg cccaacctgc
     3361 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3421 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3481 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3601 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3661 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3721 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3781 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3841 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4921 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5821 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5881 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     5941 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     6001 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
