LOCUS       sequence-442957-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Chaplin1_DR 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  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 6503)
  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-Chaplin1_DR
FEATURES             Location/Qualifiers
     source          1..6503
                     /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(2691..2709)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2692..2709)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2729..2746)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2735..2959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3005..3433
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3092..3111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3302..3323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3442..3462)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3447..3776
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3627..3762
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3689..3708
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3942..4063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3979..3998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4033..4052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4112..4128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4112..4128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4125..4147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4136..4152
                     /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(4160..4166,4167..4184,4185..4190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4205..4226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4343..4360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4514..5102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4594..4613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5273..6064,6065..6133))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     5896..5915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6134..6238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6313..6332)
                     /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 ggacgcccgc ggcaaaaagc gcaaaatcac ggaggagaac cgtggcttca acgtgagttg
      781 gaccgaaagt ttcgccttcg tggccaatgc ggaaggcctc ccagagtgtc tgctctgtag
      841 cgaaaagctg agcaacaaca agaagagcaa tgtggagcgc cattttcaag gccgtcacgc
      901 gaccttcgcg gccgaatatc cagtgggtag cgaacgcaaa agcgccatcg cgctgctggt
      961 ggaaaaactc gaagaacgca aaaataaatt caaaaaatgg atcgcgagtc caaatagcac
     1021 gaccgcggcg agttttgtgg cgacgcgcga gatcatcaag cgcggcaaac cgtttaccga
     1081 cggcgactac atgaaggaga gctttatcaa tattagcgaa catctcttca gcgactttaa
     1141 aaacaaaacg gagatcatcc agaagattaa agacatgccg ctgagcgcga agaccgttaa
     1201 agaacgtgcc atcaagatgg cgggcaacat cacggagcaa cagattaagg acattaacag
     1261 cgccccggcg tacagcattg cgtgcgacga aagctgcgat gttaccgaca tcgaacagac
     1321 cgcgctgctg tgccgttatg tgaatagcga tggcccacaa gaagaaatca tcaaactgat
     1381 cccgctcaag ggccaaaccc gtggcgagga catctgcgaa gcggtgctga agtgtctgaa
     1441 cgaaaacggt atcaatacca accacctcat cagcgtggcc acggatggtg ccccgagtat
     1501 gcgtggtagc aagcgcggct ttgtgacgct gctgcagaag gcgctcgatc gcaatctgct
     1561 ggcgttccat tgcatcctcc accaagaagc gctctgcgcc cagaccttcc caagcgagtg
     1621 tatggtggtg atgaacctcg ttatcgaaat ggtgaacaag atcatcgcca aggcgctgaa
     1681 tcatcgccag tttcgcgcgc tgctcgacga ggtggacagc gaatacagtg atctgctgct
     1741 gcacaacaag gtgcgctggc tcagtaagga tgaggttctg cgtcgctttg ttgcgtgtct
     1801 ggagcacgtt aagacgttcc tcaagagcaa ggatctgatc tacccacagc tggaggatac
     1861 ggaatggctg gagaagctgc acttcatggt ggacatgacg agccacctca acaagctcaa
     1921 cgaaagtctg caagtgcgtg gcaatacggc gctccaaatg ctggaagccg ttctcagttt
     1981 cgaacgcaaa ctgaccgttt tcgcgcgcga tgttcaacgt ggcaccctca gtcactttcc
     2041 gagtctgcgc gagttcaaag agagtcatca agatcatacg atcaacggcg actatctgca
     2101 caacgcgatc gtggacatgc aaacggcctt cgagagccgc ttctgcgagt tccgcaaaga
     2161 gaagatgacc ctcagctttc cggtgacccc actggaaatc gatccgagtc tgctgagcac
     2221 gtttccgggt atcattcaag ccgatctgga aatggagatg gcggacatca gcgacaagga
     2281 tctgtgggtg agcaagttca aacgcctcac ggcggaactc gaggatgtta cccgtcagaa
     2341 ggcgcagctc gcgcaaagtc acaagtggag cgagatggaa ggtctgccag tgccggaaaa
     2401 gctcatctat gacacgtgga acgccctccc ggactgctat aaaaacatga aaacctacgc
     2461 cttcggcgtg ctgagcattt tcggcagtac gtatctctgc gagcagatct ttagtaacat
     2521 gaactatatc aaaagcaaat atcgtacgcg tctgacccac gagagcctcc agagttgcgt
     2581 taagatcaaa gttaccagct acatgccgga tgtggagaag ctcagcagtg acgtgcgcaa
     2641 acagaagagt cattaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     2701 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2761 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2821 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2881 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2941 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3001 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3061 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3121 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3181 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3241 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3301 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3361 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3421 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3481 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3541 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3601 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3661 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3721 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3781 tccccgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4081 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5281 gcttaatcag tgaggcacct atctcagcga tctgtctctt tcgttcatcc atagttgcct
     5341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5401 caataatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5581 ccatcgctac aggcatcgtg gtatcacgct cgtcgtttgg tatggcttca ttcagctccg
     5641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gcgcaaaaaa gcggttagct
     5701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc cgtgttatca ctcatggtta
     5761 tggcagcact acataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6241 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6301 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6361 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6421 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6481 tgtacgggcc agatatacgc gtt
//
