LOCUS       sequence-437706-        6121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding GFP tagged human Tipin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6121)
  AUTHORS   Prorok P, Wolf E, Cardoso MC
  TITLE     Timeless-Tipin interactions with MCM and RPA mediate DNA replication
            stress response.
  JOURNAL   Front Cell Dev Biol. 2024 Feb 29;12:1346534. doi: 
            10.3389/fcell.2024.1346534. eCollection 2024.
  PUBMED    38487270
REFERENCE   2  (bases 1 to 6121)
  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: "Front Cell 
            Dev Biol."; date: "2024-02-29"; volume: "12:1346534. doi"; pages: " 
            10.3389/fcell.2024.1346534. eCollection 2024"
COMMENT     Sequence Label: pEGFP-hTipin (pc4732)
FEATURES             Location/Qualifiers
     source          1..6121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      866..1001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1333..1352
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1351..1406
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1452..1471
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          1603..2175
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             complement(join(3133..3843,3844..3846,3847..3849))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3133..3843,3844..3846,3847..3849))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3175..3196)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3524..3543
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3783..3804
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3846..3855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3868..3888)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     5'UTR           3924..3963
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     primer_bind     complement(4032..4056)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4053..4256)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4082..4102)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4257..4560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4583..4602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4584..4602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4721..4740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4840..4862
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4900..4918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4986..5090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5091..5159,5160..5951)
                     /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(5309..5328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaatt agtcagcaac
      841 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
      961 tgagctattc cagaagtagt gaagaggctt ttttggaggc ctaggctttt gcaaaaagct
     1021 ccgggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     1081 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     1141 aaaacaaaat ataaaaaaaa tctaacctca agtcaaggct tttctatgga ataaggaatg
     1201 gacagcaggg ggctgtttca tatactgatg acctctttat agccaacctt tgttcatggc
     1261 agccagcata tgggcatatg ttgccaaact ctaaaccaaa tactcattct gatgttttaa
     1321 atgatttgcc ctcccatatg tccttccgag tgagagacac aaaaaattcc aacacactat
     1381 tgcaatgaaa ataaatttcc tttattagcc agaagtcaga tgctcaaggg gcttcatgat
     1441 gtccccataa tttttggcag agggaaaaag atctcagtgg tatttgtgag ccagggcatt
     1501 ggccacacca gccaccacct tctgataggc agcctgcacc tgaggagtga attctttgcc
     1561 aaaatgatga gacagcacaa caaccagcac gttgcccagg agctgtagga aagagaagaa
     1621 ggcatgaaca tggttagcag aggggcccgg tttggactca gagtatttta tcctcatctc
     1681 aaacagtgta tatcattgta accataaaga gaaaggcagg atgatgacca gggtgtagtt
     1741 gtttctacca ataagaatat ttccacgcca gccagaattt atatgcagaa atattctacc
     1801 ttatcattta attataacaa ttgttctcta aaactgtgct gaagtacaat ataatatacc
     1861 ctgattgcct tgaaaaaaaa gtgattagag aaagtactta caatctgaca aataaacaaa
     1921 agtgaattta aaaattcgtt acaaatgcaa gctaaagttt aacgaaaaag ttacagaaaa
     1981 tgaaaagaaa ataagaggag acaatggttg tcaacagagt agaaagtgaa agaaacaaaa
     2041 ttatcatgag ggtccatggt gatacaaggg acatcttccc attctaaaca acaccctgaa
     2101 aactttgccc cctccatata acatgaattt tacaatagcg aaaaagaaag aacaatcaag
     2161 ggtccccaaa ctcaccctga agttctcagc tctagatcat ctagcttcag taatatttct
     2221 ggatgtagca tcaagttgct gttgcacatt tttaaaagac tggtccagca gtgtttcctc
     2281 ttcatttaaa gtattggcaa tagcatcatt acatggattg tccagaatgt cttcgtttaa
     2341 tccatttgac tcctcctttt gatcctcatc agtattaacc tcttcaaccg tgtgtgccct
     2401 gggtgtattc attaacatat catttcctag ggtctgacta ttactcagca gctttgcctg
     2461 ccttctttcc aaggccagtt gtttatttct ctcaattctt tgttgttgct cttctgttag
     2521 gcttctactt aactcagaag caaacatctc actttcagat aagtttgtca gaaagggatc
     2581 taattcagta gaagtgacat catgttcatt attctccgca acttcatcat tattgctaac
     2641 aaaatcttca tgtaaaatag ggagatcaag tcgaattcgt tttaaacagg tctgaacttc
     2701 ctttttactt cccaggtatt caactctgtc aataaaatcc tcaaactgca gtttagggaa
     2761 tagcctatgt gcccagtgct ccatgtgtct gattagcatc ttcaagtctt cagcctcatg
     2821 acctttacct ttgaattttg ccttatcaaa tacatgcctt aaggctggaa gtcctctctc
     2881 tgaaattaat ctctgagcat ccagcttggg tatatttctt ttaactgttc tctttggagg
     2941 tacaggaaca ggtgctccat ttcctgactc ttcatcaggc tcagttcctt caccatcttg
     3001 tctctctgga gaggctggag gtgggaaagg aggaaaagtt tcatcttcta catgctcata
     3061 atctggtagg tcaatcacgc cattctcctg tggttctagc atcgaagctt gagctcgaga
     3121 tctgagtccg gacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     3181 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     3241 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     3301 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     3361 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     3421 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     3481 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     3541 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     3601 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     3661 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     3721 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     3781 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     3841 gctcaccatg gtggcgaccg gtaccagtac tttgcgcttc ttcttcgggg atccccacga
     3901 agccatacgc gcttctacaa ggcgctggcc gaagaggtgc gggagtttca cgccaccaag
     3961 atcccccgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
     4021 cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac ctcccaccgt
     4081 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     4141 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     4201 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     4261 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     4321 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     4381 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     4441 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     4501 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     4561 gggcgagctc gaattccggt ctccctatag tgagtcgtat taatttcgat aagccagacc
     4621 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     4681 ggagtcgcat aagggagagc gtcgaatggt gcactctcag tacaatctgc tctgatgccg
     4741 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4801 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4861 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4921 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5641 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5701 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5761 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5821 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5881 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5941 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6001 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6061 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6121 c
//
