LOCUS       sequence-415364-        9172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human POLG gene on pcDNA3.1 backbone. POLG gene is cloned 
            with BamHI and XbaI restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9172)
  TITLE     Mammalian expression plasmid with POLG gene
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9172)
  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            "Unpublished"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: POLG-pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..9172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4672..4713
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4723..4740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4763..4780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4769..4993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5039..5467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5126..5145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5336..5357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5476..5496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5481..5810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5661..5796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5877..6671
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5931..5950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6541..6560
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6847..6968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6884..6903)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6938..6957
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7017..7033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7017..7033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7030..7052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7041..7057
                     /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)."
     promoter        complement(join(7065..7071,7072..7089,7090..7095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7110..7131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7248..7265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7419..8007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7499..7518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8178..8969,8970..9038))
                     /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     8801..8820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9039..9143)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gttaagcttg gtaccgagct cggatccatg agccgcctgc tctggaggaa ggtggccggc
      961 gccaccgtcg ggccagggcc ggttccagct ccggggcgct gggtctccag ctccgtcccc
     1021 gcgtccgacc ccagcgacgg gcagcggcgg cggcagcagc agcagcagca gcagcagcag
     1081 cagcaacagc agcctcagca gccgcaagtg ctatcctcgg agggcgggca gctgcggcac
     1141 aacccattgg acatccagat gctctcgaga gggctgcacg agcaaatctt cgggcaagga
     1201 ggggagatgc ctggcgaggc cgcggtgcgc cgcagcgtcg agcacctgca gaagcacggg
     1261 ctctgggggc agccagccgt gcccttgccc gacgtggagc tgcgcctgcc gcccctctac
     1321 ggggacaacc tggaccagca cttccgcctc ctggcccaga agcagagcct gccctacctg
     1381 gaggcggcca acttgctgtt gcaggcccag ctgcccccga agcccccggc ttgggcctgg
     1441 gcggagggct ggacccggta cggccccgag ggggaggccg tacccgtggc catccccgag
     1501 gagcgggccc tggtgttcga cgtggaggtc tgcttggcag agggaacttg ccccacattg
     1561 gcggtggcca tatccccctc ggcctggtat tcctggtgca gccagcggct ggtggaagag
     1621 cgttactctt ggaccagcca gctgtcgccg gctgacctca tccccctgga ggtccctact
     1681 ggtgccagca gccccaccca gagagactgg caggagcagt tagtggtggg gcacaatgtt
     1741 tcctttgacc gagctcatat cagggagcag tacctgatcc agggttcccg catgcgtttc
     1801 ctggacacca tgagcatgca catggccatc tcagggctaa gcagcttcca gcgcagtctg
     1861 tggatagcag ccaagcaggg caaacacaag gtccagcccc ccacaaagca aggccagaag
     1921 tcccagagga aagccagaag aggcccagcg atctcatcct gggactggct ggacatcagc
     1981 agtgtcaaca gtctggcaga ggtgcacaga ctttatgtag gggggcctcc cttagagaag
     2041 gagcctcgag aactgtttgt gaagggcacc atgaaggaca ttcgtgagaa cttccaggac
     2101 ctgatgcagt actgtgccca ggacgtgtgg gccacccatg aggttttcca gcagcagcta
     2161 ccgctcttct tggagaggtg tccccaccca gtgactctgg ccggcatgct ggagatgggt
     2221 gtctcctacc tgcctgtcaa ccagaactgg gagcgttacc tggcagaggc acagggcact
     2281 tatgaggagc tccagcggga gatgaagaag tcgttgatgg atctggccaa tgatgcctgc
     2341 cagctgctct caggagagag gtacaaagaa gacccctggc tctgggacct ggagtgggac
     2401 ctgcaagaat ttaagcagaa gaaagctaag aaggtgaaga aggaaccagc cacagccagc
     2461 aagttgccca tcgagggggc tggggcccct ggtgatccca tggatcagga agacctcggc
     2521 ccctgcagtg aggaggagga gtttcaacaa gatgtcatgg cccgcgcctg cttgcagaag
     2581 ctgaagggga ccacagagct cctgcccaag cggccccagc accttcctgg acaccctgga
     2641 tggtaccgga agctctgccc ccggctagac gaccctgcat ggaccccggg ccccagcctc
     2701 ctcagcctgc agatgcgggt cacacctaaa ctcatggcac ttacctggga tggcttccct
     2761 ctgcactact cagagcgtca tggctggggc tacttggtgc ctgggcggcg ggacaacctg
     2821 gccaagctgc cgacaggtac caccctggag tcagctgggg tggtctgccc ctacagagcc
     2881 atcgagtccc tgtacaggaa gcactgtctc gaacagggga agcagcagct gatgccccag
     2941 gaggccggcc tggcggagga gttcctgctc actgacaata gtgccatatg gcaaacggta
     3001 gaagaactgg attacttaga agtggaggct gaggccaaga tggagaactt gcgagctgca
     3061 gtgccaggtc aacccctagc tctgactgcc cgtggtggcc ccaaggacac ccagcccagc
     3121 tatcaccatg gcaatggacc ttacaacgac gtggacatcc ctggctgctg gtttttcaag
     3181 ctgcctcaca aggatggtaa tagctgtaat gtgggaagcc cctttgccaa ggacttcctg
     3241 cccaagatgg aggatggcac cctgcaggct ggcccaggag gtgccagtgg gccccgtgct
     3301 ctggaaatca acaaaatgat ttctttctgg aggaacgccc ataaacgtat cagctcccag
     3361 atggtggtgt ggctgcccag gtcagctctg ccccgtgctg tgatcaggca ccccgactat
     3421 gatgaggaag gcctctatgg ggccatcctg ccccaagtgg tgactgccgg caccatcact
     3481 cgccgggctg tggagcccac atggctcacc gccagcaatg cccggcctga ccgagtaggc
     3541 agtgagttga aagccatggt gcaggcccca cctggctaca cccttgtggg tgctgatgtg
     3601 gactcccaag agctgtggat tgcagctgtg cttggagacg cccactttgc cggcatgcat
     3661 ggctgcacag cctttgggtg gatgacactg cagggcagga agagcagggg cactgatcta
     3721 cacagtaaga cagccactac tgtgggcatc agccgtgagc atgccaaaat cttcaactac
     3781 ggccgcatct atggtgctgg gcagcccttt gctgagcgct tactaatgca gtttaaccac
     3841 cggctcacac agcaggaggc agctgagaag gcccagcaga tgtacgctgc caccaagggc
     3901 ctccgctggt atcggctgtc ggatgagggc gagtggctgg tgagggagtt gaacctccca
     3961 gtggacagga ctgagggtgg ctggatttcc ctgcaggatc tgcgcaaggt ccagagagaa
     4021 actgcaagga agtcacagtg gaagaagtgg gaggtggttg ctgaacgggc atggaagggg
     4081 ggcacagagt cagaaatgtt caataagctt gagagcattg ctacgtctga cataccacgt
     4141 accccggtgc tgggctgctg catcagccga gccctggagc cctcggctgt ccaggaagag
     4201 tttatgacca gccgtgtgaa ttgggtggta cagagctctg ctgttgacta cttacacctc
     4261 atgcttgtgg ccatgaagtg gctgtttgaa gagtttgcca tagatgggcg cttctgcatc
     4321 agcatccatg acgaggttcg ctacctggtg cgggaggagg accgctaccg cgctgccctg
     4381 gccttgcaga tcaccaacct cttgaccagg tgcatgtttg cctacaagct gggtctgaat
     4441 gacttgcccc agtcagtcgc ctttttcagt gcagtcgata ttgaccggtg cctcaggaag
     4501 gaagtgacca tggattgtaa aaccccttcc aacccaactg ggatggaaag gagatacggg
     4561 attccccagg gtgaagcgct ggatatttac cagataattg aactcaccaa aggctccttg
     4621 gaaaaacgaa gccagcctgg accattgaat ctagagggcc cgcggttcga aggtaagcct
     4681 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat
     4741 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4801 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4861 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4921 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4981 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     5041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5101 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5221 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5341 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5401 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5461 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5641 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5701 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5761 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5821 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5881 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5941 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6001 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6061 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6121 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6181 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6241 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6301 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6361 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6421 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6481 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6541 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6601 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6661 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     6721 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6781 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6841 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6901 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6961 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7021 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7081 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7141 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cg
//
