LOCUS       sequence-490353-        6599 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 6599)
  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 6599)
  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: pNP2314 [Cloning backbone, Gluc/Cluc Inversion...
FEATURES             Location/Qualifiers
     source          1..6599
                     /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"
     CDS             complement(1436..1990)
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1987..1996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2076..2180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2181..2249,2250..3041)
                     /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(2399..2418)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3212..3800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3701..3720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3954..3971
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3974..4181
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4212..5870)
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     primer_bind     complement(5941..5960)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5942..5960)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6005..6208)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6034..6054)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6209..6588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
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 aaccccaccc
     1261 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     1321 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     1381 gatggctggc aactagaagg cacagtcgag gctgatcagc gagctctagc tcgaggtcac
     1441 caccggcccc cttgatcttg tccacctggc cctggatctt gctggcaaag gtcgcacagc
     1501 gttgcggcag ccacttcttg agcaggtcag aacactgcac gttggcaagc cctttgaggc
     1561 agccagttgt gcagtccaca cacagatcga cctgtgcgat gaactgctcc atgggctcca
     1621 agtccttgaa cccaggaatc tcaggaatgt cgacgatcgc ctcgcctatg ccgccctgtg
     1681 cggactcttt gtcgccttcg taggtgtggc agcgtcctgg gatgaacttc ttcatcttgg
     1741 gcgtgcactt gatgtgggac aggcagatca gacagcccct ggtgcagcca gctttccggg
     1801 cattggcttc catctctttg agcacctcca gcggcagctt cttgccgggc aacttcccgc
     1861 ggtcagcatc gagatccgtg gtcgcgaagt tgctggccac ggccacgatg ttgaagtctt
     1921 cgttgttctc ggtgggcttg gcctcggcca cagcgatgca gatcagggca aacagaactt
     1981 tgactcccat ggtggcagtg gagacgcgtc tctgctccca ccatgggata gtgataagga
     2041 tagtgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2101 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2161 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2221 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2281 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2341 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2401 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2461 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2521 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcta
     2581 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2641 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2701 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2761 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2821 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2881 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2941 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3001 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3061 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3121 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3181 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3241 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3301 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3361 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3421 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3481 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3541 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3601 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3661 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3721 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3781 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3841 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3901 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3961 cagtgagcga ggatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     4021 ctgcccaacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     4081 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     4141 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgagc
     4201 tctagctcga gtttgcattc atctggtact tctagggtgt ctccatgctt tatgtagcat
     4261 tctttcttga actcccatgc gtggtcacag aaaccctgtt gtcccctcag gcaatactcg
     4321 tacatgcatc gttcgacacg gtcaggcttg tggcacacgt tacatttctg atcaagatca
     4381 ctttgaccgg cgaagagact attgcagagt cgttcagctt caggtttctg ttcttcggtg
     4441 catcccggtg ggttgggggt cagatcacag gctccttcag catcaaaaga atcatcactg
     4501 aaatcctggt tgtagttacc gcaaataccg caagtcttac catcgaatgg atctctaata
     4561 tgtacgacga gcagttgctt gaagttgaac ttgaccacca gagcttcagg taggatggct
     4621 gtagtcagta tgtcaccatc ttgccagtag atggaagtgt tctgagagct gtacgggacg
     4681 gacacggctt ctcctccaac cagaatctgt tttccatcaa caatgagttc tactacagtc
     4741 gattgtttcc tgattcgtac tttctctact tcagtgtaag agtcaacatt cctgtgtgaa
     4801 accttcacat cccaagtgtt ccagaaacag tcggcggcca taagaatctc cttgcaggga
     4861 ccctggaatt ggtatcttgc tttgtcaaat gtatcgtaga aagtatgtcc agacaaaacg
     4921 caggttcctc tagtttcggg agcagcgcac gtttccctgt agtcaagcag cacgtgtgag
     4981 gctcgctcgt cgccacccat acagcgggcg aaggcgcagg agatggtgta gtagtagaag
     5041 ttgatggggt tgcggcagct gtccttgtac ggctccagaa gacctttgca gtatgcaacg
     5101 tcatcaggat ttccatagag tggacaaccg tcaaactcct ggttgatctt aggctgaata
     5161 gcgagttgtt ctggatctga agtgaagtct gtatcttcca tcattttaag atctccacag
     5221 aggccagaga tcattccttt gtttgctgtg tctggggcga ttcttacaga tcttcctccg
     5281 aggatgtcga tcacgatcag tttgaagaac tcaatgacgg tgatgttgaa gcctggcatc
     5341 tcaacaacag cgatggtgac ctcgccgatg gtgtacgggt tggcgatgat agggtcagct
     5401 ccaccgttta cagtgatggg attctcagta gcttgagcga tgtcaatgat gtctccagcc
     5461 acttccagtc ttgtcttggt cagcacagcc cccttggttc catccaggtt ctccagggtg
     5521 atggacacct tccagtcgcc gcccttggtt ccttgaccca acacgtatgt accaggttcc
     5581 tggaactgga atctctttcc atagaatgtt ctaaaccatc ctgcggcctc tactctgcat
     5641 tcaattacat actgacacat tcgacaacat gtttttcctg gtttattttc acacagtcca
     5701 tctgatagta tgtctctcgt gcaggtgcca cagctgctat caatacattc tccttcttta
     5761 gcttcacagg aagttggaac tgtgtttggt ggatcaggtt cgtaaggaca gtcctggcaa
     5821 tgaacagtgg cgcagtagac taatgcaacg gcaagaatta aggtcttcat ggtggcggcg
     5881 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgggtca
     5941 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
     6001 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     6061 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     6121 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     6181 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     6241 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     6301 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     6361 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     6421 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     6481 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     6541 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcag cggaagagc
//
