LOCUS       sequence-436473-        8743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NEIL1-mCherry in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8743)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 8743)
  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: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: NEIL1-mCherry
FEATURES             Location/Qualifiers
     source          1..8743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1642..1945
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1946..2149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3400..4110
                     /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(3545..3563)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3813..3832
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        4138..4637
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4173..4196)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4570..4589
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4658..5257
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4658..4677)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5154..5174
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5442..5874
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5977..5999)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6135..6155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6140..6469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6320..6455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6491..6510
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6591..6608)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6762..7350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6842..6861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7521..8312,8313..8381))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8144..8163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8382..8486)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8554..8572
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8672..8743,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc atagttatta atagtaatca attacggggt cattagttca
     1621 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1681 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1741 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1801 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1861 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1921 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1981 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2041 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2101 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2161 ccgtcagatc cgctagccgc caccatgcct gagggccccg agctgcacct ggccagccag
     2221 tttgtgaatg aggcctgcag ggcgctggtg ttcggcggct gcgtggagaa gtcctctgtc
     2281 agccgcaacc ctgaggtgcc ctttgagagc agtgcctacc gcatctcagc ttcagcccgc
     2341 ggcaaggagc tgcgcctgat actgagccct ctgcctgggg cccagcccca acaggagcca
     2401 ctggccctgg tcttccgctt cggcatgtcc ggctcttttc agctggtgcc ccgcgaggag
     2461 ctgccacgcc atgcccacct gcgcttttac acggccccgc ctggcccccg gctcgcccta
     2521 tgtttcgtgg acatccgccg gttcggccgc tgggaccttg ggggaaagtg gcagccgggc
     2581 cgcgggccct gtgtcttgca ggagtaccag cagttcaggg agaatgtgct acgaaaccta
     2641 gcggataagg cctttgaccg gcccatctgc gaggccctcc tggaccagag gttcttcaat
     2701 ggcattggca actatctgcg ggcagagatc ctgtaccggc tgaagatccc cccctttgag
     2761 aaggcccgct cggtcctgga ggccctgcag cagcacaggc cgagcccgga gctgaccctg
     2821 agccagaaga taaggaccaa gctgcagaat ccagacctgc tggagctatg tcactcagtg
     2881 cccaaggaag tggtccagtt ggggggcaaa ggctacgggt cagagagcgg ggaggaggac
     2941 tttgctgcct ttcgagcctg gctgcgctgc tatggcatgc caggcatgag ctccctgcag
     3001 gaccggcatg gccgtaccat ctggttccag ggggatcctg gaccgttggc acccaaaggg
     3061 cgcaagtccc gcaaaaagaa atccaaggcc acacagctga gtcctgagga cagagtggag
     3121 gacgctttgc ctccaagcaa ggccccttcc aggacacgaa gggcaaagag agaccttcct
     3181 aagaggactg caacccagcg gcctgagggg accagcctcc agcaggaccc agaagctccc
     3241 acagtgccca agaaggggag gaggaagggg cgacaggcag cctctggcca ctgcagaccc
     3301 cggaaggtca aggctgacat cccatccttg gaaccagagg ggacctcagc ctctggcgga
     3361 ggatcgtcga cggtaccgcg ggcccgggat ccaccggtca tggtgagcaa gggcgaggag
     3421 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3481 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3541 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3601 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3661 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3721 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3781 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3841 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3901 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3961 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     4021 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     4081 accggcggca tggacgagct gtacaagtga gcggccgcga ctctagataa ttctaccggg
     4141 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     4201 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     4261 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     4321 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     4381 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     4441 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     4501 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     4561 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     4621 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt
     4681 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4741 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4801 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4861 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4921 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4981 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5041 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5101 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5161 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5221 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
     5281 cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgatcgagat atcagtggtc
     5341 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     5401 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     5461 aggtttggca agctactagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5521 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc gatcgaccct
     5581 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5641 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5701 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     5761 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5821 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     5881 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     5941 tggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6001 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6061 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     6121 gtagatccgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttactgg
     6481 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6541 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     6601 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6661 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6721 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6781 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6841 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6901 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6961 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7021 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7081 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7141 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7201 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7261 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7321 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7381 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7441 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7501 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7561 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7621 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7681 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7741 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7801 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     7861 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7921 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7981 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8041 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8101 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     8161 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8221 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8281 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8341 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8401 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8461 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8521 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8581 cttcaagaat tagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat
     8641 attggctcat gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag
     8701 taatcaatta cggggtcatt agttcatagc ccatatatgg agt
//
