LOCUS       sequence-207984-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GUS and Hygromycin resistance gene from SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  TITLE     pSV40 expression constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6463)
  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            "Unpublished"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: pSV40-GUS-P2A-Hygro
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..197
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      48..183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             233..2041
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             2051..2107
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2108..3133
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2108..3133
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3205..3326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3216..3235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3270..3289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3576..3596
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3748..3765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3919..4507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3999..4018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4678..5469,5470..5538))
                     /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     5301..5320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5539..5643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5670..6125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5757..5776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5967..5988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6256..6304
                     /note="synthetic polyadenylation signal"
     misc_feature    6318..6409
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6358..6377
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
       61 ctccgcccag ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag
      121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      181 gcctaggctt ttgcaaaaag cttggcattc cggtactgtt ggtaaagcca ccatgttacg
      241 tcctgtagaa accccaaccc gtgaaatcaa aaaactcgac ggcctgtggg cattcagtct
      301 ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa agcgcgttac aagaaagccg
      361 ggcaattgct gtgccaggca gttttaacga tcagttcgcc gatgcagata ttcgtaatta
      421 tgcgggcaac gtctggtatc agcgcgaagt ctttataccg aaaggttggg caggccagcg
      481 tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa gtgtgggtca ataatcagga
      541 agtgatggag catcagggcg gctatacgcc atttgaagcc gatgtcacgc cgtatgttat
      601 tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac gaactgaact ggcagactat
      661 cccgccggga atggtgatta ccgacgaaaa cggcaagaaa aagcagtctt acttccatga
      721 tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc cgaacacctg
      781 ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg cgtctgttga
      841 ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg atcaacaggt
      901 ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc acctctggca
      961 accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga cagagtgtga
     1021 tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggcgaac agttcctgat
     1081 taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg acttgcgtgg
     1141 caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact ggattggggc
     1201 caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact gggcagatga
     1261 acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct ctttaggcat
     1321 tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag tcaacgggga
     1381 aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca aaaaccaccc
     1441 aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag gtgcacggga
     1501 atatttcgcg ccactggcgg aagcaacgcg taaactcgac ccgacgcgtc cgatcacctg
     1561 cgtcaatgta atgttctgcg acgctcacac cgataccatc agcgatctct ttgatgtgct
     1621 gtgcctgaac cgttattacg gatggtatgt ccaaagcggc gatttggaaa cggcagagaa
     1681 ggtactggaa aaagaacttc tggcctggca ggagaaactg catcagccga ttatcatcac
     1741 cgaatacggc gtggatacgt tagccgggct gcactcaatg tacaccgaca tgtggagtga
     1801 agagtatcag tgtgcatggc tggatatgta tcaccgcgtc tttgatcgcg tcagcgccgt
     1861 cgtcggtgaa caggtatgga atttcgccga ttttgcgacc tcgcaaggca tattgcgcgt
     1921 tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa ccgaagtcgg cggcttttct
     1981 gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa ccgcagcagg gaggcaaaca
     2041 aggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc
     2101 tggaccgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     2161 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     2221 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     2281 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     2341 tgacattggg gaatttagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     2401 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     2461 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     2521 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     2581 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     2641 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     2701 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     2761 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     2821 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     2881 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     2941 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     3001 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     3061 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     3121 ggcaaaggaa tagttctaga caaacaccat acaacaagaa agggccctag agtcggggcg
     3181 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3241 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3301 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3361 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3421 tcgataagga tctgaacgat ggagcggaga atgggcggaa ctgggcggag ttaggggcgg
     3481 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     3541 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     3601 cggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     3661 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     3721 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4321 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5641 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6121 aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6181 ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt
     6241 acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     6301 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     6361 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgataggta
     6421 ccgagctctt acgcgtgcta gcccgggctc gagatctgcg atc
//
