LOCUS       sequence-442903-        5594 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Tc1-15_Xt transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  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 5594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Tc1-15_Xt
FEATURES             Location/Qualifiers
     source          1..5594
                     /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     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1782..1800)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1783..1800)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1820..1837)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1826..2050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2096..2524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2183..2202)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2393..2414
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2533..2553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2538..2867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2718..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2780..2799
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3033..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3070..3089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3124..3143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3203..3219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3203..3219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3216..3238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3227..3243
                     /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(3251..3257,3258..3275,3276..3281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3296..3317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3434..3451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3605..4193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3685..3704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4364..5155,5156..5224))
                     /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     4987..5006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5225..5329)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5404..5423)
                     /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 gcgtcgcagc ttacagccta cccaaatggc gcaagtggtg cagttaatcc aggatggcac
      781 cagcatgcgt gcggttgcgc gtcgttttgc agttagcgtg agcgttgttt cacgtgcgtg
      841 gcgccgttat caagagactg gccagtatat acgccgtcgt ggtggtggtc gtcgtcgcgc
      901 gactacccaa caacaggatc gctacttacg tttatgcgcg cgccgtaacc gtcgtagcac
      961 agcacgtgcg ctgcagaacg atttacaacg cgccaccaac gttcatgtta gcgcccagac
     1021 tgtgcgtaac cgcttacatg agggcggtat gcgtgcacgt cgtcctcaag ttggtgtggt
     1081 tttaaccgcg caacatcgtg caggccgctt agcgtttgca cgcgaacatc aggattggca
     1141 gattcgtcat tggcgcccgg tgctgtttac cgatgagagc cgctttaccc tgagcacctg
     1201 tgatcgtcgc gatcgtgttt ggcgtcgtcg tggtgaacgt agcgcagcat gcaacattct
     1261 gcagcatgat cgctttggca gcggtagcgt gatggtgtgg ggtggtattt cactgggtgg
     1321 tcgcaccgcg ttacatgttt tagcgcgcgg cagcttaacc gcaattcgct atcgcgacga
     1381 aattctgcgc cctctggttc gtccttatgc aggtgcagtt ggccctggtt tcctgctgat
     1441 gcaggataac gcccgccctc atgttgcagg tgtgtgccaa caattcctgc aggatgaagg
     1501 cattgacgcg atggactggc ctgcacgttc acctgacctg aaccctattg agcatatctg
     1561 ggatattatg agccgcagca ttcaccagcg tcacgttgca cctcagaccg tgcaagaact
     1621 ggtggacgcg ttagtgcaag tgtgggagga gattccgcag gaaaccattc gccacctgat
     1681 tcgctcaatg cctcgtcgtt gtcgcgaggt tattcaagca cgtggcggcc acacccatta
     1741 ttgatgagcg gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat
     1801 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1861 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1921 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1981 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2041 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2161 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2221 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2281 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2401 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2461 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2521 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgaaa
     2881 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2941 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3001 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3181 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4021 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4141 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4201 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4261 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4321 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4441 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4561 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4801 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4861 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5041 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5221 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5281 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5341 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5401 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5461 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     5521 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     5581 cagatatacg cgtt
//
