LOCUS       sequence-444758-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Tigger2 transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6452)
  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 6452)
  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-Tigger2
FEATURES             Location/Qualifiers
     source          1..6452
                     /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(2640..2658)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2641..2658)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2678..2695)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2684..2908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2954..3382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3041..3060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3251..3272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3391..3411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3396..3725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3576..3711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3638..3657
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3891..4012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3928..3947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3982..4001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4061..4077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4061..4077)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4074..4096)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4085..4101
                     /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(4109..4115,4116..4133,4134..4139))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4154..4175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4292..4309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4463..5051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4543..4562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5222..6013,6014..6082))
                     /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     5845..5864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6083..6187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6262..6281)
                     /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 gggccctatt cgtagcgcga cctcagatgc aggtagcgcc cctaaaaaac agcgcaaggt
      781 gatgaccctg caggaaaaag tggaactgct ggacatgtat cgtcgtttac gtagcgcagc
      841 agcagttgca cgccacttta aaattaacga aagcagcgtg cgcaccatcg tgaaaaagga
      901 gaaggaaatt cgcgaagccg ttgcagcagc aactcctgcg ggtgcgaaaa ccttacattt
      961 cctgcgcaac accttcctga gccgcataga aaacgccgcg ttcatgtggg tgcaggattg
     1021 ctacaaaaaa ggcattccga tcgacagcaa catgattcgc gagaaggcca agagcctgta
     1081 cgataacctg aaacagaaag aaggcgaagg cagcaaagcg ggcgagttca acgcaagcaa
     1141 aggctggttt gacaactttc gcaaacgctt cggcctgaag aacgtgaaaa ttaccggcga
     1201 agcggcatca gcagaccaag aagcggcaga cgaatttccg gatgcgatta agaagatcat
     1261 cgaggagaaa ggctatctgc ctgaacaggt gttcaacgcg gacgaaagcg ccttattttg
     1321 gcgcaagaag atgcctcagc gcaccttcat tagcaaagaa gagaagcgcg cgcctggttt
     1381 taaagcgggt cgtgatcgtc tgaccctgct gttttgtgcg aacgcggtgg gtttcatgat
     1441 tcgcaccgcg ctgatttata aagccgccaa ccctcgtgcg ttaaagggca aagacaagca
     1501 tcagctgccg gtgttctggc tgtataacaa gaaagcgtgg accacccgca ccttatttct
     1561 ggactggttc catcgctgct ttgtgcctga agtgcgcaaa tatctggcga gcaagggcct
     1621 gccgtttaaa gtgctgctga tcctggataa tgcgcctggt catccggaac cgcatgaatt
     1681 taacaccgaa ggcgtggagg ttgtgtatct gccgcctaac accacctcac tgattcagcc
     1741 gctggatcag ggtgtgatcc gcacctttaa agcccattat acccgctata gcatggagcg
     1801 cattgtgaac gcgatggaag aaaacccgga ccgcgagaac attatgaagg tgtggaagga
     1861 ctataccatc gaagatgcga tcgtggtgat cgaaaaagcc gtgaaggcca tcaaaccgga
     1921 aaccattaac agctgctggc gcaaactgtg tcctgacgtg gtgcatgatt ttaccggctt
     1981 caccaccgag cctattaagg aaatcatgaa ggagattgtg gacatggcga aaaaagttgg
     2041 cggcgagggc tttcaagata tggacctggg cgagattcag gagctgatcg acaccacccc
     2101 tgaggaactg accgaagacg atctgatgga gatgagcgcg agcgaacctg tgcctgatga
     2161 cgaagaggag gacgtggaag aagccgtgcc ggaaaacaaa ctgaccctgg acaacttagc
     2221 ggagggcttt cgcttattca aaaccgcctt cgacttcttc tatgacatgg accctagcat
     2281 gatccgcgcg ctgaaactga aacagaccgt ggaagaaggc ttagtgccgt atcgcaacat
     2341 ttttcgcgag atgaagaaac agaagagcca gaccgagatt accatgtact tccataaagt
     2401 gaccccgagc gttcctgcaa gccctgcatc accttcaacc agcagcacta gcagcgcgtc
     2461 agcaacctta cgtcaacagg accagcctct gctgttcctg ttactgctga gcctgctgaa
     2521 cgtgaaaacc atgcgcatga aaaccttcat gatgatccac ttccatctga tgaacagcaa
     2581 gtatatcttc agcagcctgt aatgagcggc cgctcgagca tgcatctaga gggccctatt
     2641 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3721 gcaaaaagct ccccgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3781 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3841 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3901 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3961 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4021 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4081 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4141 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4201 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4261 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4321 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4381 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4441 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4621 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4861 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5221 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5281 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5341 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5401 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5461 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5521 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5581 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5641 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5701 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5761 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5821 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5881 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5941 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6001 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6061 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6121 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6181 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6241 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6301 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6361 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6421 gcttcgcgat gtacgggcca gatatacgcg tt
//
