LOCUS       sequence-378106-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor for targeted integration of mNG_hSLBP(18-108) probe to the 
            AAVS1 safe harbor locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  TITLE     S-phase specific probe
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AAVS1_Puro_H2BK_mNG_hSLBP(18-108)
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    320..1123
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1130..1155
                     /label=SA
                     /note="splice acceptor site"
     CDS             1179..1232
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1236..1832
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1729..1749
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1848..1969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1859..1878)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1913..1932
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2860..3567
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     misc_feature    4515..5351
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(5428..5445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5599..6187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5679..5698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6358..7149,7150..7218))
                     /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     6981..7000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7219..7323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7391..7409
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagaa ttcgcccttt gctttctctg accagcattc tctcccctgg gcctgtgccg
      361 ctttctgtct gcagcttgtg gcctgggtca cctctacggc tggcccagat ccttccctgc
      421 cgcctccttc aggttccgtc ttcctccact ccctcttccc cttgctctct gctgtgttgc
      481 tgcccaagga tgctctttcc ggagcacttc cttctcggcg ctgcaccacg tgatgtcctc
      541 tgagcggatc ctccccgtgt ctgggtcctc tccgggcatc tctcctccct cacccaaccc
      601 catgccgtct tcactcgctg ggttcccttt tccttctcct tctggggcct gtgccatctc
      661 tcgtttctta ggatggcctt ctccgacgga tgtctccctt gcgtcccgcc tccccttctt
      721 gtaggcctgc atcatcaccg tttttctgga caaccccaaa gtaccccgtc tccctggctt
      781 tagccacctc tccatcctct tgctttcttt gcctggacac cccgttctcc tgtggattcg
      841 ggtcacctct cactcctttc atttgggcag ctcccctacc ccccttacct ctctagtctg
      901 tgctagctct tccagccccc tgtcatggca tcttccaggg gtccgagagc tcagctagtc
      961 ttcttcctcc aacccgggcc cctatgtcca cttcaggaca gcatgtttgc tgcctccagg
     1021 gatcctgtgt ccccgagctg ggaccacctt atattcccag ggccggttaa tgtggctctg
     1081 gttctgggta cttttatctg tcccctccac cccacagtgg ggcaagcttc tgacctcttc
     1141 tcttcctccc acagggcctc gagagatctg gcggcggaga gggcagagga agtcttctaa
     1201 catgcggtga cgtggaggag aatcccggcc ctagcaccga gtacaagccc acggtgcgcc
     1261 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     1321 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     1381 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     1441 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     1501 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     1561 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     1621 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     1681 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     1741 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     1801 cctggtgcat gacccgcaag cccggtgcct gatctagaca gacatgataa gatacattga
     1861 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1921 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1981 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aatgcaggac
     2041 tagtgaaaca aacatgactg tgtccagctt tctctaagac agtgtagaat ggctctgaaa
     2101 agagcctttg ttttccagtt ttcacaacct cgctccttat ttggccttat ggtggctctc
     2161 agtcttctta ggcagcagca cggcctggat attgggcaag acaccaccct gcgcgatggt
     2221 gactttgccc agcagcttgt tgagctcctc gtcgttgcgg atggccagtt ggaggtgacg
     2281 cgggatgata cgggtcttct tgttgtcgcg ggccgcattg ccagccagct ccaggatctc
     2341 agcggtcagg tactccagca ccgcagccag gtacactggc gcgccggctc caacccgctc
     2401 ggcataatta cccttgcgga gcaggcggtg cactcggccc acggggaact gaagcccagc
     2461 ccgagaagag cgggtcttgg ccttggcgcg agctttaccg ccttgcttgc cacgtccaga
     2521 catactggtc acaacttact gaaagtagcc aacgaggaaa ggcagcgcct tttataaaca
     2581 ctattgggcg cgaaaaagaa gatatgtcat tggttagggt tgtagcttag ttttaaccaa
     2641 tggaagtacg tcttttggat ttgcagattg ggttgattgg gtataactaa cctctgacgt
     2701 caccctaata accaccaatc agatacagga ctttcactct tcaccttatt tgcatgagag
     2761 gtcctatata aaaacgactc ctggtacctt gctcccatta cttcccgttt tctcgatctg
     2821 ctgctcgtct caggctcgta gttcgccttc aactgtacca tggtgagcaa gggcgaggag
     2881 gataacatgg cctctctccc agcgacacat gagttacaca tctttggctc catcaacggt
     2941 gtggactttg acatggtggg tcagggcacc ggcaatccaa atgatggtta tgaggagtta
     3001 aacctgaagt ccaccaaggg tgacctccag ttctccccct ggattctggt ccctcatatc
     3061 gggtatggct tccatcagta cctgccctac cctgacggga tgtcgccttt ccaggccgct
     3121 atggtagatg gctccggata ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc
     3181 cttactgtta actaccgcta cacctacgag ggaagccaca tcaaaggaga ggcccaggtg
     3241 aaggggactg gtttccctgc tgacggtcct gtgatgacca actcgctgac cgctgcggac
     3301 tggtgcaggt cgaagaagac ttaccccaac gacaaaacca tcatcagtac ctttaagtgg
     3361 agttacacca ctggaaatgg caagcgctac cggagcactg cgcggaccac ctacaccttt
     3421 gccaagccaa tggcggctaa ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg
     3481 gagctcaagc actccaagac cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat
     3541 gtgatgggca tggacgagct gtacaaggga tatccatcac actggcggcc gctcgaggcc
     3601 agcccgccgt cccccgcgcg atggagcctg ggacggaagc gcagagccga cggcaggcgc
     3661 tggaggcccg aagacgccga ggaggcagag caccgcggcg ccgagcgcag acccgagagc
     3721 tttaccactc ctgaaggccc taaaccccgt tccagatgct ctgactgggc aagtgcagtt
     3781 gaagaagatg aaatgaggac cagagttaac aaagaaatgg caagatataa aaggaaactc
     3841 ctcatcaatg actttggaag agagagaaaa tgataatgta caaaagcgtc ttaaagccca
     3901 accccaaagg ctcttttaag agccacttaa attatcgata ttagagctgt aaacacgtgt
     3961 tcctaatcca ggcttaaatt cgggcgatct ttttaaaggg attatcatga tcttatcaca
     4021 ctgagtaatg catgcagtgc ttgtaatcga gtctggctaa ggagaagcca catacacttt
     4081 gttagtactg agtgaggaat atggcgccta aaaaattagg gatgtgcagg aaacttgtgt
     4141 taacagaaag tgcttcctgg ctcttggcgc gaaaatggat acttccgttg tgtcgagtgc
     4201 tgtgacttcc tgttagaact tgttgaaagc ctattgtgtc acgtgtactt tccaccatgt
     4261 aatggcgttc taacgtgagt ttatatatta gtaaatttgg ttcccgatgg cgttccagta
     4321 cgtatggctt tcgctcgagc agttacctta tttaacctct tctaggctgt acccagctct
     4381 gccttgcttt gcttccccct caccccctga ttccatctct gcagcccacg ggacatcggg
     4441 aagatgccag accaccaggc aggatagtct gcaagggtaa ggggaggagc acagcgtcga
     4501 cagtactaag ctttactagg gacaggattg gtgacagaaa agccccatcc ttaggcctcc
     4561 tccttcctag tctcctgata ttgggtctaa cccccacctc ctgttaggca gattccttat
     4621 ctggtgacac acccccattt cctggagcca tctctctcct tgccagaacc tctaaggttt
     4681 gcttacgatg gagccagaga ggatcctggg agggagagct tggcaggggg tgggagggaa
     4741 gggggggatg cgtgacctgc ccggttctca gtggccaccc tgcgctaccc tctcccagaa
     4801 cctgagctgc tctgacgcgg ctgtctggtg cgtttcactg atcctggtgc tgcagcttcc
     4861 ttacacttcc caagaggaga agcagtttgg aaaaacaaaa tcagaataag ttggtcctga
     4921 gttctaactt tggctcttca cctttctagt ccccaattta tattgttcct ccgtgcgtca
     4981 gttttacctg tgagataagg ccagtagcca gccccgtcct ggcagggctg tggtgaggag
     5041 gggggtgtcc gtgtggaaaa ctccctttgt gagaatggtg cgtcctaggt gttcaccagg
     5101 tcgtggccgc ctctactccc tttctctttc tccatccttc tttccttaaa gagtccccag
     5161 tgctatctgg gacatattcc tccgcccaga gcagggtccc gcttccctaa ggccctgctc
     5221 tgggcttctg ggtttgagtc cttggcaagc ccaggagagg cgctcaggct tccctgtccc
     5281 ccttcctcgt ccaccatctc atgcccctgg ctctcctgcc ccttccctac aggggttcct
     5341 ggctctgctc taagggcgaa ttctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6001 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6961 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7321 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7381 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
