LOCUS       sequence-443167-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of PROTOP transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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-PROTOP
FEATURES             Location/Qualifiers
     source          1..7163
                     /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(3351..3369)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3352..3369)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3389..3406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3395..3619
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3665..4093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3752..3771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3962..3983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4102..4122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4107..4436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4287..4422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4349..4368
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4639..4658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4693..4712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4772..4788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4772..4788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4785..4807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4796..4812
                     /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(4820..4826,4827..4844,4845..4850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4865..4886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5003..5020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5174..5762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5254..5273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5933..6724,6725..6793))
                     /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     6556..6575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6794..6898)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6973..6992)
                     /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 ggcccatctg aacaacctga tggatatggc caagaagaag cgccgctata tcagcggcaa
      781 caaatgcagc ctgagccatt gtcgtaaaag ccgccgcgat tttcctgcgc tgaagctgtt
      841 ccgttttccg gaacgcgatc cgaccatgtt aattcgctgg gcggaaaaat gccagtttac
      901 cgaagatttc gtggcgagca ccagcagctt atttctgtgc caggatcact tcagcccgga
      961 tgatattggc gtgaagtacc tgaagaaagg cgccattcct gatcgcaacc tgatcaaata
     1021 taagagctgc aacaacgccg atatcaacct gaatgcggtg ggtagcccgc ctcgtaaacg
     1081 tcatcgtagc cagagccctg ttgaagtgtg tcgccagtgc cataagaacg ccaaaaccct
     1141 gaaagtgttc cagaagaagt acttcgcgtt taagaagctg agcgatgaac gccagatccg
     1201 cattaagatg ctgcgcaagg agaacagcaa cctgaaacgc aagctggtgc gcctggaaag
     1261 caaaaccgaa aagaacatct acagcgagat cgataaaatc aacctgaccg gcaacgagaa
     1321 aattctggcc aaaatgctgc tgaaagaaaa gaaaggcacc aacacccgct ggaacgattg
     1381 cgaaaaactg ttcgcgcaga gcatttacta tcgcagcacc agcacctata ccttcctgcg
     1441 cgatagcctg aaactgaact ttccgagccc tagcagcctg cagaaatgga acagcattaa
     1501 gaagctgcag ccgggcgata acgaatgcct gtatagcgcc ctgaaagagg ccatcaagga
     1561 aatgaacgcg agcgataaag aatgcattct ggcgtgcgat gaggtggcga tcaaaaaaaa
     1621 cctgacctat aacgtgagcg tggatatcat tgacggcatt gaacacctgc tggatcgcag
     1681 caacaaaatc ggcagccaca tttgcgtgtt tgtggtgcgc ggcattctga aaaagtggaa
     1741 gtttatcctg aactatttcg tggcggaaac caacattaaa ggcgactgcc tgaaaagcct
     1801 gatctacaag aacatcatca tcgccgaaac cattggcttt aaagtgcgcg gcgtggtgta
     1861 tgatcagggc ggcaacaacc gtaaatgcac cagcctgctg gaagtgacca acgagaaacc
     1921 gtacttcacc ctgaacaaca agaagtacta catgttctac gatatcccgc atctgtttaa
     1981 gagcgtgcgc aacaactttc tgaaagccaa ctttgaaacc cctgacggcc tggtggattt
     2041 tgacgtgatt cgcgaagtgt atgaactgga tcatggcagc gttacccgca tgaccaaact
     2101 gacccgcagc catgttaatc cgacccgctt tgaattaatg cgcgtgtgct tagcaaccca
     2161 gaccctgagc catactgttg cggcggcgat taaaacctgc aaccagaaca aacagctgca
     2221 ccgcaacagc tcagaagttg cagcgagcac cgcagcattt gtgcagaaag tgaacgacta
     2281 ctttgattgc ctgaacagcc gtgtgctgac cgacaaaaac ccgatgaaat gcgcgctgca
     2341 ggtgaataac ggcgtgtgga acaaactgaa agagatgcag gaatatctga agagcgtgaa
     2401 atatcacggc aacaagattt attgcgtgga tggcctgatc caaaccaccg aagcgatttt
     2461 tggcctggtg gaggatctgt ttaaagacca taccgaccac ttcttttttc tgaccagccg
     2521 cgtgaaccaa gatccgctgg aaaacatttt tgcgtgcgtg cgcgcgaaag gtggtaattg
     2581 ccgcaaccct agcgtgaacg agttcaacat catcatcgcg aaactgatca gcctgcacat
     2641 ctttaaattc agccagaaga gcaactgcga aagcgatgac gatgtgatgc tgccgattga
     2701 gttcgacagc attatttatc agccgttcat cgagaagaaa gaaatccagc agcaggaata
     2761 tagcgtgagc ttcagcaaaa ttgtgcagga taacgagcgc tactttgacc agaacatcga
     2821 taactttctg tgcaacgatg tgccgatcga attaaccagc agccgctatt tcgtgggcta
     2881 tattgcgaaa ggcagcagct gtgacaaatg ccgcagcgtg attctgaaag aaaccgaaca
     2941 cctgaccgcg cctagcgaac tgttcatcca tgaaaagaac tacagcattg agagcgattt
     3001 cggcaaactg aaagccccga gcgatctgtt ttttaacatc tgcaagatcc acattaaggc
     3061 gttcgaaaac atcttcaaga acaacaaaaa gcagatgtgc atcaagaagt ttattgtgga
     3121 acagtgcatc aaatgcacca acgaaagcag cgcctttagc ctgtggtttt acgaaaacaa
     3181 cgagtgctat gcgcatcgca ccgatctgct gaacaaactg attaaggtgc tgctgtttaa
     3241 acactgtaag tggaccgtga ttgccgatcg ccagaagaaa caggcgaagc tgagcattct
     3301 gagccacaaa tgatgagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     3361 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3421 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3481 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3541 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3601 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3661 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3721 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3781 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3841 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3901 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3961 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4021 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4081 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4201 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4261 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4321 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4381 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4441 tccccgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4501 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4561 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4741 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4801 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4861 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4921 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4981 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5101 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5161 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5221 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5281 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5341 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5401 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5461 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5521 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5581 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5641 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5701 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5761 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5821 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5881 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5941 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6001 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6061 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6121 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6181 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6241 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6301 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6361 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6421 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6481 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6541 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6601 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6661 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6721 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6781 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6841 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6901 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6961 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7021 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7081 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7141 tgtacgggcc agatatacgc gtt
//
