LOCUS       sequence-442906-        5603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Tc1-16_Xt transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  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 5603)
  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-Tc1-16_Xt
FEATURES             Location/Qualifiers
     source          1..5603
                     /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(1791..1809)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1792..1809)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1829..1846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1835..2059
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2105..2533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2192..2211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2402..2423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2542..2562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2547..2876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2727..2862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2789..2808
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3079..3098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3133..3152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3212..3228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3212..3228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3225..3247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3236..3252
                     /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(3260..3266,3267..3284,3285..3290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3305..3326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3443..3460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3614..4202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3694..3713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4373..5164,5165..5233))
                     /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     4996..5015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5234..5338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5413..5432)
                     /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 ggacaaccgt aaacgccgcc gcgaactgtc agaagatctg cgcaccaaga ttgtggaaaa
      781 atatcagcag agccagggct ataaaagcat tagccgcgac ttagacttac cgctgagcac
      841 cgtgcgcaac atcattaaga agtttgcgac ccatggcacc gtggcaaact tacctggtcg
      901 tggccgcaaa cgcaaaattg atgaacgcct gcagcgtcgt atagtgcgca tggtggacaa
      961 acagcctcag accagcagca aagagatcca ggccgtgtta caagcgcaag gtgcgtcagt
     1021 tagcgcacgt accattcgcc gccatctgaa cgaaatgaaa cgctatggcc gtcgtcctcg
     1081 ccgtacccct ttattaaccc agcgccataa aaaagcgcgc ctgcagtttg cgaagatgta
     1141 tctgagcaag ccgcagagct tttgggaaaa cgtgctgtgg accgatgaaa ccaagatcga
     1201 actgtttggc aaagcgcacc atagcaccgt gtatcgcaaa cgcaacgaag cgtacaagga
     1261 aaaaaacacc gtgcctaccg tgaaatatgg cggcggcagc atgatgtttt ggggctgctt
     1321 tgcagcgtca ggtaccggtt gtctggaatg cgtgcagggc attatgaaaa gcgaggacta
     1381 tcagcgtatt ctgggccgca ccgttgaacc tagcgtgcgt aaattaggcc tgcgtcctcg
     1441 ctcatgggtg tttcagcagg ataacgatcc taaacatacc agcaaaagca cccagaaatg
     1501 gatggcgacc aagcgttggc gtgttctgaa atggcctgcc atgagccctg atctgaaccc
     1561 gattgaacat ctgtggcgcg acctgaaaat tgcggtgggc aaacgccgtc ctagcaacaa
     1621 acgcgatctg gagcagtttg cgaaagaaga gtggagcaaa attccgggcg aacgctgcaa
     1681 aaagctgatc gacggctatc gcaaacgcct gattagcgtg attttcagca aaggctgcgc
     1741 caccaagtat tgatgagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     1801 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1861 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1921 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1981 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2041 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2101 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2521 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2641 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2701 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2761 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2821 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2881 tccccgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     2941 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3001 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3181 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3361 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5341 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     5401 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     5461 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     5521 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     5581 tgtacgggcc agatatacgc gtt
//
