LOCUS       sequence-490392-        5702 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 5702)
  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 5702)
  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: pNP3254 [pNP2922 - ISCro4 Enhanced DBL (DBL3, ...
FEATURES             Location/Qualifiers
     source          1..5702
                     /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           2257..2324
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     promoter        2391..3569
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2622..3560
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3517..3537
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3576..3585
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4381..4390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4581..4601
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4602..4622
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4623..4643
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4653..4709
                     /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(4716..4718,4719..5432)
                     /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(4716..4718,4719..5429)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4758..4779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5019..5038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5366..5387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5457..5474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5463..5670
                     /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 caccgcgttc
     2161 tccacaagtg acaaaccgga cagtatcatg gaccggtttt cccggtaatc cgcattcaca
     2221 aggctggtct cacttgtgga gaacgtttta ttttttggcc ggcatggtcc cagcctcctc
     2281 gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggactttttt tgttctagtc
     2341 cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc ggctccggtg
     2401 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     2461 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     2521 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     2581 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
     2641 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
     2701 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
     2761 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
     2821 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
     2881 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
     2941 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
     3001 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     3061 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     3121 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     3181 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     3241 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     3301 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     3361 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     3421 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     3481 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     3541 gtttttttct tccatttcag gtgtcgtgac gtacggccac catggaacag gagctgcact
     3601 tcatcggcat cgacgtgtcc aaagcaaagc tggacgtgga cgtgctgcga cctgacggca
     3661 gacacagaag caagaaattc gctaataccc ctaagggaca cgacgagctg ctgagatggc
     3721 tgagcggcca cagagtggcc ccagctcaca tctgcatgga agctacaagc acctacatgg
     3781 aggatgtggc cgcccacctg agcgacgccg gctacaccgt gtccgtgatc aaccctgccc
     3841 tgggcaaggc ctttgcccaa agcgagggac tgcgaagcaa gacagatgct gttgatgcca
     3901 gaatgctggc cgagttctgt cggcagaagc ggcctccagc ctgggaggcc cctcaccctg
     3961 tggaacgggc cctgagggcc ctcgtgctgc gccaccagag cctgaccgat atgcacaccc
     4021 aggagctgaa ccgactggaa accgcccggg aagtgcagcg gcctagcatt gacgcccatc
     4081 tgctgtggct gcacgccgag ctgaaaagaa tcgagaaaca gatcaaggac ctgacagatg
     4141 atgaccccga catgaagcac cggcggaagc tgctggaaag catccccggc atcggagaaa
     4201 agaccagcgc cgtcctgctc gcttatacag gcctgaagga aagattcacc cacgccagac
     4261 agttcgctgc ttttgccggc ctgacaccta gaagatacga gagcggcagc tctgtgaacc
     4321 gggcctctag aatgagcaag gccggacatg cctctctgcg gagagccctg tacatgcctg
     4381 ccatggtggc cgtgtctaag accgagtggg gcagagcctt ccgggacaga ctggccggca
     4441 acggcaagaa gggcaaagtg atcatcggcg ccatgatgag aaagctggcc caggtggcct
     4501 acggcgtgct gaagtccggc gtccccttcg acgccagcag acacaacccc gtggctgctg
     4561 ctagcggagg gggtggaagc ccaaaaaaga agagaaaggt acctaagaaa aagcggaagg
     4621 tccccaagaa gaagaggaaa gttggaagcg gtgccacgaa cttctctctg ttaaagcaag
     4681 caggcgacgt agaagaaaac ccaggtcccg gatccgtgag caagggcgag gagctgttca
     4741 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4801 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4861 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4921 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4981 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5041 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5101 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5161 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     5221 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5281 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     5341 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     5401 tcactctcgg catggacgag ctgtacaagt aactcgagct agagctcgct gatcagcctc
     5461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5641 ttgggaagac aatagcaggc atgctgggga cgacgtcagg tggcactttt cggggaaatg
     5701 tg
//
