LOCUS       sequence-318096-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a fragment of mouse Pola1 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  AUTHORS   Li Z, Hua X, Serra-Cardona A, Xu X, Gan S, Zhou H, Yang WS, Chen CL,
            Xu RM, Zhang Z
  TITLE     DNA polymerase alpha interacts with H3-H4 and facilitates the 
            transfer of parental histones to lagging strands.
  JOURNAL   Sci Adv. 2020 Aug 26;6(35):eabb5820. doi: 10.1126/sciadv.abb5820. 
            eCollection 2020 Aug.
  PUBMED    32923642
REFERENCE   2  (bases 1 to 5543)
  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: "Sci Adv."; 
            date: "2020-08-26"; volume: "6(35):eabb5820. doi"; pages: " 
            10.1126/sciadv.abb5820. eCollection 2020 Aug"
COMMENT     Sequence Label: pGST-POLA1-N0
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        244..321
                     /gene="lacI"
                     /label=lacI promoter
     CDS             322..1404
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(341..360)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1453..1458,1459..1476,1477..1483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1491..1507
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1515..1531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1515..1531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1527..2054
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1547..1564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1547..1563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1556..1578)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2156..2161,2162..2177,2178..2184)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2192..2208
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2231..2884
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2842..2864
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2897..2917
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3566..3588)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3733..3751)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3819..3923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3924..3992,3993..4784)
                     /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     complement(4142..4161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4955..5543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5444..5463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat
      241 tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
      301 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
      361 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
      421 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
      481 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
      541 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
      601 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
      661 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
      721 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
      781 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
      841 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
      901 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
      961 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1021 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1081 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     1141 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     1201 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     1261 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     1321 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1381 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     1441 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1501 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
     1561 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1621 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1681 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
     1741 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     1801 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
     1861 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     1921 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     1981 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     2041 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     2101 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     2161 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
     2221 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     2281 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     2341 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     2401 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     2461 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     2521 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     2581 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     2641 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     2701 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     2761 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     2821 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     2881 aaaaatcgat acgaccgaaa acctgtattt tcagggcgcc atgatggcgc ccatgcacga
     2941 agaggactgt aaactggagg caagcgctgt gtcagattca gggagttttg cagcttcccg
     3001 agctcggcga gaaaagaaat caaagaaagg acgtcaagaa gctttagaga gactgaaaaa
     3061 ggctaaagct ggggagaagt ataaatacga aatgggatag gctatgtgga agatggcaga
     3121 gagatttttg atgatgacct tgaggatgat gcacttgaca cctgtggaaa aggaagtgat
     3181 ggcaaagcac acagaaagga caggaaggat gtaaagaaac cttcagtgac aaaaccaaac
     3241 aatatcaagg caatgttcat tgctagtgcc gggaagaaaa ctacagataa agcagtagac
     3301 ttgtccaagg atgatctgct aggtgacatt ctacaggatc taaacactga gacagctcaa
     3361 ataactccac cacctgtatt gataccaaag aaaaaaagat ctactggggc tttgctcaac
     3421 cctttttgag atccggaatt caaaggccta cgtcgacgag ctcaactagt gcggccgctt
     3481 tcgaatctag agcctgcagt ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg
     3541 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3601 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3661 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     3721 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3781 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3841 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3901 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3961 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4021 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4081 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4141 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4201 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4261 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4321 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4441 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     4501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5101 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5521 caggggggcg gagcctatgg aaa
//
