LOCUS       sequence-490364-        4140 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Cloning backbone for measuring bridge recombinase mediated 
            inversion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4140)
  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 4140)
  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: pNP2322 [Cloning backbone, mCherry Inversion
            A...
FEATURES             Location/Qualifiers
     source          1..4140
                     /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    1198..1405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1394..1411
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1436..2146)
                     /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(1714..1733)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1983..2001
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2143..2152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2232..2336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2337..2405,2406..3197)
                     /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(2555..2574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3368..3956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3857..3876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4110..4127
                     /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 gagacgcgtc tccgccgtcc
     1201 ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     1261 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     1321 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     1381 gatggctggc aactagaagg cacagtcgag gctgatcagc gagctctagc tcgagttact
     1441 tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca
     1501 cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc
     1561 cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc
     1621 cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt
     1681 acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg
     1741 aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg
     1801 agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga
     1861 agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg
     1921 ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac
     1981 ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc
     2041 cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga
     2101 actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg gcagtggaga
     2161 cgcgtctctg ctcccaccat gggatagtga taaggatagt gacgtcaggt ggcacttttc
     2221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2341 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2401 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2461 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2521 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2581 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2641 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2701 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2761 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2821 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2881 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2941 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3001 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggatctcgcg
     3061 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3121 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3481 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4021 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4081 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
//
