LOCUS       sequence-444807-        7103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Zaphod2 transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7103)
  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 7103)
  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-Zaphod2
FEATURES             Location/Qualifiers
     source          1..7103
                     /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"
     promoter        complement(3291..3309)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3292..3309)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3329..3346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3335..3559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3605..4033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3692..3711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3902..3923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4042..4062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4047..4376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4227..4362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4289..4308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4542..4663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4579..4598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4633..4652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4712..4728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4725..4747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4736..4752
                     /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(4760..4766,4767..4784,4785..4790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4805..4826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4943..4960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5114..5702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5194..5213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5873..6664,6665..6733))
                     /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     6496..6515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6734..6838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6913..6932)
                     /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 gagcgagaag aacttcaaaa aactgagcgg cagcgagaac cgcaaatgga aagcgattcg
      781 cgaagagaac gaacgcaaac tgtttggcag cctgaacaaa ttcctggaaa agatcgatgg
      841 catgccggaa aagaaagaaa aaaccagcga gctgctggtt agcgcagaat gcagcaccga
      901 cacctcactg atggagcagg atgtgaactt accgagcacc agcatgagca ccatcaccga
      961 tgaatttcag gaaaagctgg aggaaaaaaa gagcgaggag agcattaaac ctaacgtgct
     1021 gagcgcggat cctggtactt ggccttgcaa cctgaacatt aagcagcgcg ataccctggt
     1081 ggaaaactat cctccgcagg tgcgcaactt caactttcct aaagatagca ccggccgcaa
     1141 gttcagcgaa acctactata cccgcatcct gcctaacggc gaaaaaatta cccgcagctg
     1201 gctgttatat agcaccagca aggatagcgt gttttgcctg tactgcaaac tgtttggcga
     1261 tggcaaaaac cagctgaaaa acgaaaacgg ctgcaaagat tggcagcatc tgagccatat
     1321 cctgagcaaa catgaagaga gcgagatgca tatcaacaac agcgtgaaat acagcaaact
     1381 gaagagcgac ctgaagaagt acatcaccat tgatgccgcg gaacagcgcc tgtatgaaaa
     1441 cgaagagaaa tattggcgcg cggtggtgga gcgcattatt tacatgatcc agtttctggc
     1501 gcgccaatgc ttagcgtttc gtggcagcag caaaaagctg tacgagcgcg acaacggcaa
     1561 ctttctgcat ctggcggaaa ccattgcgaa attcgataac attatggcgg agcatctgtg
     1621 ccgtattcag accagcaccc cgaacaacat tacccattat ctgggcgacg agattcagaa
     1681 ccagatcatc aagattattg ccgatgcgat gaacgagatc attctgaaca aagtgcgcaa
     1741 cgcgaaatat tatagcatca tcctggatac caccccggat gtgtcacatg tggagcagat
     1801 caccattatt gtgcgctttg tggattttca gaacgccaac attagcatct gcgaacattt
     1861 tggcggcttt tgcccggtgg atgatattac cggcgaaggc ctgtatacct ttctggtgga
     1921 ttacctgcag aagctgaacc tggataaaaa caacctgcgc ggccagggtt atgataacgc
     1981 caccaacatg catggcaaac ataacggcct gcagcagaaa attctggaaa agtatagccg
     2041 cgccttctat attccgtgca gcgcgcatag cttaaacctg gtggtgaacg atgcggcgaa
     2101 gattagcttc gataccaccg agtttttcta cattgtgcag gaactgtaca actattttag
     2161 cgcgagcacc aaacgttgga gcgtgctgaa aacccacctg ccgaacatta acctgaaacc
     2221 tctgagcaac acccgttggg aaagccgcgt tgatgcgatc atgccgctga aaacccagtt
     2281 agaagcggtg tgcaacgcgc tgctggaaat tgtggacgac atcaacaaag atattgacag
     2341 ccgcatgcag gcgaaaggca ttctgagcaa aatcagcacc ttcaagttca tttgcagcgt
     2401 gctgatttgg catgacattc tgagcaagat caacgtgatt agcaagatga tgcaggagcc
     2461 taaatttaac gtgcacaacg ccctggaaag cattaaaggc atcaagaagt tctttgagga
     2521 gaaacgcagc gatgatgtgt atgagaagta tattatcgac gcgaagcaga ttgcggaaag
     2581 cctggatatt gacgccaact tcagcaacgt ggaaaacact cgcaagcgcc gcaagaagac
     2641 ccattttgat tgcgaaagcc atgatcagcc tatttatgac ccgaagctga actttaagat
     2701 caacttcttc ttcagcattc tggacaccac cattagcagc ctgaaagaac gctttaccat
     2761 cctgaaagag tatcacggcc tgtttagcgt gctggatgat atttgcaacc tgaagaacta
     2821 taacgaacgc gaactgctgc agctgtgcaa aaacctgcag ctgaaactga ccgatgaaca
     2881 tagcaaggaa tgcgatattg acggcaccag catgtatgag gaactgaaga ccctgagctg
     2941 ctttctgaag agcagcatga cccctgttga ggtgctgcag tacatttgca gcaacaacct
     3001 gccggacacc tttccgaaca cctttgtggc cctgcgcatt tatctgaccc tgccgatcag
     3061 cgttgcaagc ggcgaacgca gctttagcaa gctgaagttc atcaagaact atctgcgcag
     3121 caccatgagc caagaacgcc tgagcaacct ggccaccatt agcattgaaa acgagattct
     3181 ggagcaggtg agcattgaaa ccgtgatcaa aaacttcatc agcatgaagg cgcgccgtgt
     3241 gaccatactg tgatgagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     3301 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3361 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3421 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3481 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3541 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3961 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4381 tccccgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4441 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4501 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4681 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6841 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6901 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6961 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7021 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7081 tgtacgggcc agatatacgc gtt
//
