LOCUS       sequence-490497-        3638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Antgra4 RFP cargo plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3638)
  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 3638)
  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"; journalName: "Nat 
            Biotechnol. 2025 Oct 2. doi: 10.1038/s41587-025-02816-4."
COMMENT     Sequence Label: pTIR_Antgra4_RFP
FEATURES             Location/Qualifiers
     source          1..3638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      514..1102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1003..1022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1317..1528
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1541..1809
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1837..1846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2583..2603)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2609..2730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2646..2665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2700..2719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2930..2948)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3016..3120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3121..3189,3190..3638,1..343)
                     /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     complement(3339..3358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
       61 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      121 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      181 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      241 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      301 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1141 ttgctgcctt aaccctttcg gccccaatgg gacaaataaa tcccaacata catttcataa
     1201 catctgctgg gcgctatctg tcccgttttt aatggaaatt gtcattagcg ggtattttaa
     1261 ggttttaaca atttggcagc tatgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     1321 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     1381 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     1441 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     1501 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     1561 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     1621 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     1681 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     1741 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     1801 ttacagatcc aagctgtgac cggcgcctac gatatcgcca ccatggccag ctccgaggat
     1861 gtcatcaaag agtttatgag atttaaggtc aagatggagg gaagcgtcaa cggacacgag
     1921 ttcgagattg agggagaagg agaaggccgg ccttacgagg gcacacaaac cgctaagctc
     1981 aaggtcacaa aaggaggacc cctccccttc tcctgggata ttctgagccc tcagttccag
     2041 tacggaagca aagcctatgt taaacaccct gccgacatcc ctgactatct gaagctctcc
     2101 ttccctgaag gcttcaagtg ggagagattc atgaacttcg aggacggagg cgtggtgaca
     2161 gtcacacaag atagcaccct ccaggacgga gagtttattt ataaggtgaa actcagagga
     2221 accaacttcc cctccgatgg ccctgtcatg caaaaaaaaa caatgggatg ggaagcctcc
     2281 accgagagaa tgtatcctga ggatggcgct ctgaaaggcg aaattaaaat gagactgaaa
     2341 ctcaaagacg gaggacacta cgatgccgag gtcaaaacaa cctacaaggc caagaaacaa
     2401 gtgcagctgc ctggcgccta catgactgat attaaactcg acattatcag ccataatggg
     2461 gactacacca tcgtggaaca atatgagaga gctgagggca gacatagcac aggcgctgga
     2521 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2581 tatgttgctc cttttacgct atgtggttaa cttgtttatt gcagcttata atggttacaa
     2641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2701 tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca
     2761 attagtctat cagaagctat ctggtctccc ttgagctccc actctccctg tatatatttt
     2821 ttttatttta actagtatat attgtcaaat gatctcttaa aataccataa ctcgcaaaaa
     2881 catccttaaa atcattaaaa ataaaattgg gtcccaaagg gttaaaggag cctcgtgata
     2941 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3001 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3061 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3121 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3181 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3241 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3301 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3361 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3421 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3481 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3541 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3601 gatcgttggg aaccggagct gaatgaagcc ataccaaa
//
