LOCUS       sequence-490476-        5791 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Express ISCro4 recombinase and bridge RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  AUTHORS   Perry NT, Bartie LJ, Katrekar D, Gonzalez GA, Durrant MG, Pai JJ, 
            Fanton A, Martins JQ, Hiraizumi M, Ricci-Tam C, Nishimasu H, 
            Konermann S, Hsu PD
  TITLE     Megabase-scale human genome rearrangement with programmable bridge 
            recombinases.
  JOURNAL   Science. 2025 Sep 25:eadz0276. doi: 10.1126/science.adz0276.
  PUBMED    40997214
REFERENCE   2  (bases 1 to 5791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2025 Sep 25:eadz0276. doi: 10.1126/science.adz0276."
COMMENT     Sequence Label: pNP3440 [pNP2922 - ISCro4 bRNA (TBL2, WT + DBL...
FEATURES             Location/Qualifiers
     source          1..5791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /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(324..343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1626..1645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1879..1896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1906..2154
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1906..1926
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2077..2096
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     ncRNA           2346..2413
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     promoter        2480..3658
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2711..3649
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3606..3626
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3665..3674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4470..4479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4670..4690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4691..4711
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4712..4732
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4742..4798
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(4805..4807,4808..5521)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4805..4807,4808..5518)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4847..4868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5108..5127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5455..5476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5546..5563)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5552..5759
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcacgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagaggg cctatttccc
     1921 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     1981 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     2041 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     2101 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgagtgc
     2161 agggagaacc ggccagttct ctctgccatg cggtccgcat gccgtatcag gcctcaggct
     2221 aataacctgt gacgtagatt tatgcagcgg accgccgttc tccacaagtg acaaaccgga
     2281 cagtatcatg gaccggtttt cccggtaatc cgcattcaca aggctggtct cactttttat
     2341 tttttggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca tgcttcggca
     2401 tggcgaatgg gacttttttt gttctagtcc gtttttagcg cgtgcgccaa ttctgcagac
     2461 aaatggctct agaggtaccg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     2521 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     2581 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     2641 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     2701 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
     2761 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
     2821 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
     2881 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
     2941 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
     3001 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
     3061 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
     3121 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
     3181 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
     3241 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
     3301 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
     3361 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
     3421 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     3481 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     3541 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     3601 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgacg
     3661 tacggccacc atggaacagg agctgcactt catcggcatc gacgtgtcca aagcaaagct
     3721 ggacgtggac gtgctgcgac ctgacggcag acacagaagc aagaaattcg ctaatacccc
     3781 taagggacac gacgagctgc tgagatggct gagcggccac agagtggccc cagctcacat
     3841 ctgcatggaa gctacaagca cctacatgga ggatgtggcc gcccacctga gcgacgccgg
     3901 ctacaccgtg tccgtgatca accctgccct gggcaaggcc tttgcccaaa gcgagggact
     3961 gcgaagcaag acagatgctg ttgatgccag aatgctggcc gagttctgtc ggcagaagcg
     4021 gcctccagcc tgggaggccc ctcaccctgt ggaacgggcc ctgagggccc tcgtgctgcg
     4081 ccaccagagc ctgaccgata tgcacaccca ggagctgaac agactggaaa ccgcccggga
     4141 agtgcagcgg cctagcattg acgcccatct gctgtggctg cacgccgagc tgaaaagaat
     4201 cgagaaacag atcaaggacc tgacagatga tgaccccgac atgaagcacc ggcggaagct
     4261 gctggaaagc atccccggca tcggagaaaa gaccagcgcc gtcctgctcg cttatacagg
     4321 cctgaaggaa agattcaccc acgccagaca gttcgctgct tttgccggcc tgacacctag
     4381 aagatacgag agcggcagct ctgtgaaccg ggcctctaga atgagcaagg ccggacatgc
     4441 ctctctgcgg agagccctgt acatgcctgc catggtggcc gtgtctaaga ccgagtgggg
     4501 cagagccttc cgggacagac tggccggcaa cggcaagaag ggcaaagtga tcatcggcgc
     4561 catgatgaga aagctggccc aggtggccta cggcgtgctg aagtccggcg tccccttcga
     4621 cgccagcaga cacaaccccg tggctgctgc tagcggaggg ggtggaagcc caaaaaagaa
     4681 gagaaaggta cctaagaaaa agcggaaggt ccccaagaag aagaggaaag ttggaagcgg
     4741 tgccacgaac ttctctctgt taaagcaagc aggcgacgta gaagaaaacc caggtcccgg
     4801 atccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4861 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4921 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4981 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     5041 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     5101 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     5161 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     5221 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     5281 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     5341 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     5401 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     5461 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     5521 actcgagcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     5581 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5641 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5701 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggac
     5761 gacgtcaggt ggcacttttc ggggaaatgt g
//
