LOCUS       sequence-490375-        6615 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Measure ISCro4 activity via luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  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 6615)
  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: pNP2389 [pNP2314 - ISCro4 Donor + ISCro4
            Targe...
FEATURES             Location/Qualifiers
     source          1..6615
                     /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"
     CDS             complement(1444..1998)
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1995..2004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2092..2196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2197..2265,2266..3057)
                     /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(2415..2434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3228..3816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3717..3736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3970..3987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3990..4197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4228..5886)
                     /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(5957..5976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5958..5976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6021..6224)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6050..6070)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6225..6604
                     /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 taacagtatc ttgtattatc
     1201 cgccgtcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccaa
     1261 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     1321 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     1381 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga gctctagctc
     1441 gaggtcacca ccggccccct tgatcttgtc cacctggccc tggatcttgc tggcaaaggt
     1501 cgcacagcgt tgcggcagcc acttcttgag caggtcagaa cactgcacgt tggcaagccc
     1561 tttgaggcag ccagttgtgc agtccacaca cagatcgacc tgtgcgatga actgctccat
     1621 gggctccaag tccttgaacc caggaatctc aggaatgtcg acgatcgcct cgcctatgcc
     1681 gccctgtgcg gactctttgt cgccttcgta ggtgtggcag cgtcctggga tgaacttctt
     1741 catcttgggc gtgcacttga tgtgggacag gcagatcaga cagcccctgg tgcagccagc
     1801 tttccgggca ttggcttcca tctctttgag cacctccagc ggcagcttct tgccgggcaa
     1861 cttcccgcgg tcagcatcga gatccgtggt cgcgaagttg ctggccacgg ccacgatgtt
     1921 gaagtcttcg ttgttctcgg tgggcttggc ctcggccaca gcgatgcaga tcagggcaaa
     1981 cagaactttg actcccatgg tggcagtttt tcgacgtagg cctgataagg ctcccaccat
     2041 gggatagtga taaggatagt gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2581 gataacactg cggctaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3361 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3841 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3901 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3961 accgagcgca gcgagtcagt gagcgaggat ccccagcatg cctgctattg tcttcccaat
     4021 cctccccctt gctgtcctgc ccaaccccac cccccagaat agaatgacac ctactcagac
     4081 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     4141 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     4201 aggctgatca gcgagctcta gctcgagttt gcattcatct ggtacttcta gggtgtctcc
     4261 atgctttatg tagcattctt tcttgaactc ccatgcgtgg tcacagaaac cctgttgtcc
     4321 cctcaggcaa tactcgtaca tgcatcgttc gacacggtca ggcttgtggc acacgttaca
     4381 tttctgatca agatcacttt gaccggcgaa gagactattg cagagtcgtt cagcttcagg
     4441 tttctgttct tcggtgcatc ccggtgggtt gggggtcaga tcacaggctc cttcagcatc
     4501 aaaagaatca tcactgaaat cctggttgta gttaccgcaa ataccgcaag tcttaccatc
     4561 gaatggatct ctaatatgta cgacgagcag ttgcttgaag ttgaacttga ccaccagagc
     4621 ttcaggtagg atggctgtag tcagtatgtc accatcttgc cagtagatgg aagtgttctg
     4681 agagctgtac gggacggaca cggcttctcc tccaaccaga atctgttttc catcaacaat
     4741 gagttctact acagtcgatt gtttcctgat tcgtactttc tctacttcag tgtaagagtc
     4801 aacattcctg tgtgaaacct tcacatccca agtgttccag aaacagtcgg cggccataag
     4861 aatctccttg cagggaccct ggaattggta tcttgctttg tcaaatgtat cgtagaaagt
     4921 atgtccagac aaaacgcagg ttcctctagt ttcgggagca gcgcacgttt ccctgtagtc
     4981 aagcagcacg tgtgaggctc gctcgtcgcc acccatacag cgggcgaagg cgcaggagat
     5041 ggtgtagtag tagaagttga tggggttgcg gcagctgtcc ttgtacggct ccagaagacc
     5101 tttgcagtat gcaacgtcat caggatttcc atagagtgga caaccgtcaa actcctggtt
     5161 gatcttaggc tgaatagcga gttgttctgg atctgaagtg aagtctgtat cttccatcat
     5221 tttaagatct ccacagaggc cagagatcat tcctttgttt gctgtgtctg gggcgattct
     5281 tacagatctt cctccgagga tgtcgatcac gatcagtttg aagaactcaa tgacggtgat
     5341 gttgaagcct ggcatctcaa caacagcgat ggtgacctcg ccgatggtgt acgggttggc
     5401 gatgataggg tcagctccac cgtttacagt gatgggattc tcagtagctt gagcgatgtc
     5461 aatgatgtct ccagccactt ccagtcttgt cttggtcagc acagccccct tggttccatc
     5521 caggttctcc agggtgatgg acaccttcca gtcgccgccc ttggttcctt gacccaacac
     5581 gtatgtacca ggttcctgga actggaatct ctttccatag aatgttctaa accatcctgc
     5641 ggcctctact ctgcattcaa ttacatactg acacattcga caacatgttt ttcctggttt
     5701 attttcacac agtccatctg atagtatgtc tctcgtgcag gtgccacagc tgctatcaat
     5761 acattctcct tctttagctt cacaggaagt tggaactgtg tttggtggat caggttcgta
     5821 aggacagtcc tggcaatgaa cagtggcgca gtagactaat gcaacggcaa gaattaaggt
     5881 cttcatggtg gcggcgaatt ccagcacact ggcggccgtt actagtggat ccgagctcgg
     5941 taccaagctt gggtcaccct atagtgagtc gtattaattt cgataagcca gtaagcagtg
     6001 ggttctctag ttagccagag agctctgctt atatagacct cccaccgtac acgcctaccg
     6061 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     6121 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     6181 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     6241 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6301 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6361 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     6421 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     6481 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     6541 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     6601 tgtcagcgga agagc
//
