LOCUS       sequence-490385-        4156 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Measure ISCro4 activity via mCherry expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4156)
  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 4156)
  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: pNP2390 [pNP2322 - ISCro4 Donor + ISCro4
            Targe...
FEATURES             Location/Qualifiers
     source          1..4156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     polyA_signal    1206..1413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1402..1419
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1444..2154)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1722..1741)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1991..2009
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2248..2352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2353..2421,2422..3213)
                     /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(2571..2590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3384..3972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3873..3892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4126..4143
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa taacagtatc ttgtattatc
     1201 cgccgtcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
     1261 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     1321 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     1381 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga gctctagctc
     1441 gagttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
     1501 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
     1561 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
     1621 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
     1681 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
     1741 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     1801 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
     1861 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
     1921 cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag
     1981 ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     2041 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     2101 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca ccatggtggc
     2161 agtttttcga cgtaggcctg ataaggctcc caccatggga tagtgataag gatagtgacg
     2221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2581 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2701 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2761 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2821 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2881 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2941 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3001 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3061 gagcgtggat ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3121 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3181 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3241 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3301 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3361 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3421 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3481 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3541 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3601 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3661 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3721 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3781 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3841 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3901 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3961 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4021 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4081 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4141 gaggaagcgg aagagc
//
