LOCUS       sequence-496981-        7261 bp    DNA     circular SYN 12-JAN-2026
DEFINITION  Antgra4 transposase expressing plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7261)
  AUTHORS   Ivancic D, Agudelo A, Lindstrom-Vautrin J, Jaraba-Wallace J, Gallo 
            M, Das R, Ragel A, Herrero-Vicente J, Higueras I, Billeci F, 
            Sanvicente-Garcia M, Petazzi P, Ferruz N, Sanchez-Mejias A, Guell M
  TITLE     Discovery and protein language model-guided design of hyperactive 
            transposases.
  JOURNAL   Nat Biotechnol. 2025 Oct 2. doi: 10.1038/s41587-025-02816-4.
  PUBMED    41039042
REFERENCE   2  (bases 1 to 7261)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 Oct 2. doi: 10.1038/s41587-025-02816-4."
COMMENT     Sequence Label: pCMV_PB_Antgra4
FEATURES             Location/Qualifiers
     source          1..7261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1269..1287)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1270..1287)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1307..1324)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1313..1537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1583..2011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1670..1689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1880..1901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2020..2040)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2025..2354
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2205..2340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2267..2286
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2421..3215
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2475..2494)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3085..3104
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3389..3510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3480..3499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3559..3575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3559..3575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3572..3594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3583..3599
                     /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(3607..3613,3614..3631,3632..3637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3652..3673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3790..3807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3961..4549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4041..4060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4720..5511,5512..5580))
                     /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     5343..5362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5581..5685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5760..5779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5951..6330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6331..6534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6485..6505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6579..6598
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6579..6597
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttactatagg gtgcacactc gtgatgtgtt ttaataaggt gcctaagctg agacattact
       61 ggagtagcca cccatctctg ggcaataaag ctattaaaag cgctatatca aagaacaggt
      121 gcttatttct gctgtccaag ctctacttca ataaccccgt gaagcctgag ggcgcctcta
      181 agatctatta cgtacaggag ctgctttcct gctttaaata cacgtttcaa aagtaccgct
      241 ccgactctac atcacagagc atcgacgaaa gtatggtggg ttttaaagga cgctccagcc
      301 tcaaacaata catgcctcag aaacccgtaa aaaggggcat caagctgtgg tcccgatgcg
      361 acgcgaagac aggttacact tacgacgcca acgtctactg cggcaaggac gatgtggaga
      421 ggtccggcac tctgggggag acagttgtta agaagttatg tgagactata cagaacccaa
      481 atgtggtcct ggccttcgat cgcttcttta ccagcgtgag gctgatggac agtctccagt
      541 accctgcagt gggcaccgcg atcataacac gaaaagatat gccaaagaaa tttatagaaa
      601 agcggaagat gtccagaggc gagtgtgagt tcttaagcaa tcagaccaac agtatcgcca
      661 tcaaatggca ggacaccaag caggtgatcc tgctgagcaa ctgtcatacc cccgatatga
      721 cttgcgtgaa acgtaaggat aagaccggtc agagggttga tgtgccatgc cccaccgcca
      781 tcgcctatta caatgaaatc atgggaggag tggaccttgc cgaccagatg tctggtgtgt
      841 atgatttcgg ccggaagagc tgtaagtggt ggaagaaggt gttttatcgc ctgctgatga
      901 tcgccgtggt gaattcctgg gtgatctata atgacctccg ccgtacgcag aagaaaatcc
      961 ccctgcttga attcctcttt acactttctg aggacctgat tgaagagggc cagcagcaga
     1021 cctctgtgaa gcttagaaga aatagttgcg ggcgtccctc taaacgcagt aaggtgatga
     1081 agaatgtagg cgatcacttg ccaattgaga catcaaagcg gcgacgctgt gctcgctgtg
     1141 cattacaaaa aaaagaaaga agaacaacca caatttgcgc tatgtgtgac gtgggcttat
     1201 gcaaacactg tttcgcactc taccacagta gctgataact gctcgagcat gcatctagag
     1261 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     1321 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1381 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1441 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1501 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1561 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1981 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2041 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2101 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2161 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2221 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2281 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2341 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     2401 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2461 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2521 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2581 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     2641 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2701 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     2761 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     2821 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     2881 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     2941 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3001 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3061 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3121 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3181 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3241 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3301 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3361 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3421 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3481 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3541 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5701 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     5761 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     5821 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     5881 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     5941 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     6001 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     6061 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     6121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     6181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     6241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     6301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     6361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     6421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     6481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     6541 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     6601 acccaagctt ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca
     6661 ccggtgtcta tgggctacat gtgtccatct gggttctccg accgcgtaat ttacattcct
     6721 atggtctttt actcaatgag tcggaagaag ttaacacgag cggagctgca ggatctggcc
     6781 gataacctcg aggagctgtc cagtggcgaa gaggtgcagt ccccatatga ggacagctcc
     6841 tctgaggagg atccatttca taacagcgag gaatctgatg agctgtacca gccttcagac
     6901 acctccgacg aagactccgg cgatatcgaa tcagacgcaa atgaaaacct gacccagcca
     6961 aatgacgggg tgatcagtgc cgagaactct gaccaagagg cggaaactcc tgtaaccact
     7021 ggaaacataa tctggtcctt acctacaaat acgtatgttc caagactctc aattcccaat
     7081 gaaagccagt gcatcatcca ccctagcatt gctgccaatg ctagccccct cgacatcttt
     7141 actaagatct gtccccgcag tctgttcact tttatagcag agagcaccaa ccagagaata
     7201 aagatctata aagagaataa gaaaaagcaa gccactttca ctgatgctgg cgaaattatg
     7261 a
//
