LOCUS       sequence-441973-        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Mariner-3_AMi 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  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 6362)
  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-3_AMi
FEATURES             Location/Qualifiers
     source          1..6362
                     /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(2550..2568)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2551..2568)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2588..2605)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2594..2818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2864..3292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2951..2970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3161..3182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3301..3321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3306..3635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3486..3621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3548..3567
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3801..3922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3838..3857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3892..3911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3971..3987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3971..3987)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3984..4006)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3995..4011
                     /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(4019..4025,4026..4043,4044..4049))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4064..4085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4202..4219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4373..4961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4453..4472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5132..5923,5924..5992))
                     /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     5755..5774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5993..6097)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6172..6191)
                     /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 ggccagcaac atgctgttca ccgccagcaa aagcaagggc gaaaaacgca aacgcgtggt
      781 gttaaccctg atgcagaaga ttgatatttg ccgctgcctg gagaaaggcg aaaaccgcaa
      841 cgtgctgatg aaagaattta gcgtgggcag cagcaccatt tacgacatca aggcgcagaa
      901 agaacagctg ctgaagtttg tgagcagcag cgaaaccagc aaagcggtgg agaaatgccg
      961 taccctgcac aagccgaaac tggagcagtt agacagcgtg ctgtatgagt ggtttagcct
     1021 gaagcgtagc gaaggcgcac ctgttagcgg tccgatgctg attgagaaag cgaaggactt
     1081 ctacaaacag atggacctga ccgaaccttg tgcgtttagc gcaggttggc tgagctgttt
     1141 taaactgcgc catggcatac gtcgtctgga tgtgagcggc gagatcaaaa gcgcggatca
     1201 cgaagcggcg gagaagtatt gcggcttctt taacaaactg gtggcggaac acgatctgag
     1261 ccctgatcag atttacaaca ccgatgaaac cggcctgttt tggcgttgcc tgcctaacag
     1321 caccttagcg ggtgcaggcg aatcaagcgc agcgggtttc aagcagaaca aggatcgcat
     1381 caccgtgctg acctgtgcaa atgcaggtgg cacccacaaa gtgaaactgc tggtggtggg
     1441 caaatgtaat cgtccgcgcg cgtttaaagg catcacccat ctgccggtgg tgtacaaagc
     1501 gcagagcaac gcgtggatgg ataagggcat cttctatgat tggttccatc atgtgtttgt
     1561 gccgagcgtg aaggaacact tccgcaaaat cggcaaaccg caggacagca aagcgattct
     1621 gctgctggat aactgccgtg cgcatcctcc tgaatgcgaa ctggtgagcg gcaacatctt
     1681 taccatcttt ctgccggcga acgtgacctc actgattcag cctatggatc agggcattat
     1741 ccagaacatg aaatgctact accgccgcga ttttctgcgc aaactgatta accacgaagg
     1801 caccattcag aacttccaga gcctgtacaa cattaaggac gcgattttta acgtggcgtg
     1861 cgcctggaat agcgtgaaga gcaagacctt acgtcgcgcg tggcgtaaat tatggccgag
     1921 cgtgatgttt gcggagggca gcagcgatga agaagatttc gaaggcttca acgtgcgcca
     1981 tcgcaaaaac accatcgcgc agattctgga ggtggtgaag gatgcgccgt caaccaaccc
     2041 tgtgaacaaa ctgaaggaaa gcgagattga agaatgggtg gaagcggaca aggaagtgga
     2101 gcacgtgttc agcgatgcgg agatcatcga aagcgtggtg aacccggaaa aaagcaaggt
     2161 gaccgaaaag gatagcgagg aagaggattt tagcgaggaa gagaaaatca gctgggcaag
     2221 cgcggcgagc tatattgata actttatcaa gtttgcggag cgccagagct gttatagcgc
     2281 gcaggaagtg atgcagctgc atattatcca cagcgacttt atgcagaaac gccaggcggc
     2341 atcacgtcag gcagatatcc gcgacttctt taagaaagcg agccgcgcct gtcacaaagc
     2401 aagcaccgaa agcgcgcctt caccttcacc tagcaccagc accattgatg tggacgatag
     2461 cgaggctctg caggatcctc aagtgcctct ggaaagcccg aagcgccagt aatgagcggc
     2521 cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     2581 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2641 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2701 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2761 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2821 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2881 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2941 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3001 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3061 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3121 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3181 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3241 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3301 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3481 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3541 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3601 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgaaatg accgaccaag
     3661 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3721 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3781 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3841 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3901 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     3961 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4021 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4081 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4141 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4201 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4261 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4801 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4921 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5041 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5101 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5161 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5221 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5281 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5341 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5401 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5461 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5521 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5581 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5641 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5701 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5761 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5821 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5881 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5941 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6001 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6061 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6121 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6181 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6241 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     6301 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     6361 tt
//
