LOCUS       sequence-442023-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Zaphod3 transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  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 5987)
  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-Zaphod3
FEATURES             Location/Qualifiers
     source          1..5987
                     /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      919..928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2175..2193)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2176..2193)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2213..2230)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2219..2443
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2489..2917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2576..2595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2786..2807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2926..2946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2931..3260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3111..3246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3173..3192
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3426..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3463..3482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3517..3536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3596..3612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3596..3612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3609..3631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3620..3636
                     /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(3644..3650,3651..3668,3669..3674))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3689..3710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3827..3844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4757..5548,5549..5617))
                     /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     5380..5399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5618..5722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5797..5816)
                     /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 gtgcgaagtg attcgctacg tgcacatcga agatggcaag gtggagatca aggagagctt
      781 tctgggcttt tttccgatta gcggcaaaac tgccgcggaa ctgaccgaag aaaccagcaa
      841 acagctggat agcgatggcc tggatattat tttctgccgc ggccagggtt atgataacgc
      901 ggcgaccgtt gcaggtaccc atggtggcgt tcgcgccaaa attaaagaga tcaaccctaa
      961 aagcctgttt gtgccgtgcg cgaaccatag cctgaatctg tgcggcgtgc atagctttgg
     1021 tagcgtgagc agctgcgtga cctttttcgg caccctggaa aaaaactaca gctttttcag
     1081 cgtgagcccg caccgttgga agatgctgca gaacgtgggc attaccgtga aacgcctgag
     1141 ccaaactcgt tggagcgcgc attatgaagc agttcgcgcg gtgaagacca actttgagaa
     1201 actgattagc acctttgaag tgctgtgcga tccgaaagaa aacgtggata cccgtggcag
     1261 cgcccagatt ttactgagcg cggtgtgcga tttcagcttc ctgagctatc tgttcttttg
     1321 gtgcgaagtg ctggacgaag tgaaccagac ccagaaatat ctgcagaccg cgggcattag
     1381 cttagaacag tgcaccgtga agcatcaggc gctgaaactg tttctggaag accgccgcac
     1441 cgaaatagtg gaaaaggcca ttaactatgc gaccaccaag tgcaaggaaa tggacatcta
     1501 catcgaaaag cgcatcaaat ttcgccgtcg catgcctggt gaaaccacca aagatgccgg
     1561 tctgacctta ccggaggaaa ttaaacgcgc catgttcgaa tgcctggatc gctttcatca
     1621 ggagctggat acccgcagca aagcgatgga tcagatcatg agcatgtttg cgatcattca
     1681 gccgtttagc ctgatttttg ccgaggaaga aaaactgcgc aaattcctgc cgaacatcat
     1741 cgaaatctac gatgagttca gcggcgaaga cattctggtg gaaattttcc gcctgcgccg
     1801 ccatttaaaa gcagcgcgca ttgatccgga agaaaccaaa acctggaccg tgctgcagtt
     1861 cctggaattt attgtgaagt gggactttta cgaaagcctg ccgaacttaa gcttatgcct
     1921 gcgcctgttt ctgaccattt gcattagcgt ggcgagctgc gaacgcaact ttagcaaact
     1981 gaagctgatt aaaagcgtgc tgcgcagcac tatgagcgaa gaccgcttaa ccaacctggc
     2041 cattctgagc attgagcacg agtacgccaa aaagatcaac tttgacgagg tgatcgataa
     2101 atttgccgaa gtgaaggcgc gcaaacagaa gctgtaatga gcggccgctc gagcatgcat
     2161 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     2221 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2281 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2341 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2401 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2461 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2521 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2581 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2641 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2701 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2761 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2821 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2881 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2941 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3121 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3181 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3241 gaggcctagg cttttgcaaa aagctccccg aaatgaccga ccaagcgacg cccaacctgc
     3301 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3361 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3421 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3541 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3601 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3661 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3721 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3781 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5041 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5341 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5761 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5821 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     5881 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     5941 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
