LOCUS       sequence-207986-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry and Hygromycin resistance gene from SV40 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  TITLE     pSV40 expression constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5455)
  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            "Unpublished"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-mCherry-P2A-Hygro
FEATURES             Location/Qualifiers
     source          1..5455
                     /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"
     regulatory      227..236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             233..940
                     /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(378..396)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     646..665
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             950..1006
                     /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             1007..2032
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1007..2032
                     /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    2125..2246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2136..2155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2190..2209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2568..2588
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2740..2757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2911..3499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2991..3010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3670..4461,4462..4530))
                     /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     4293..4312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4531..4635)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4662..5117
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4749..4768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4959..4980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5248..5296
                     /note="synthetic polyadenylation signal"
     misc_feature    5310..5401
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5350..5369
                     /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 ccatggtgag
      241 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
      301 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
      361 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
      421 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
      481 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
      541 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
      601 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
      661 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
      721 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
      781 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
      841 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
      901 gggccgccac tccaccggcg gcatggacga gctgtacaag ggatccggcg caacaaactt
      961 ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatga aaaagcctga
     1021 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     1081 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     1141 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     1201 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aatttagcga
     1261 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     1321 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     1381 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     1441 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     1501 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     1561 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     1621 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     1681 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     1741 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     1801 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     1861 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     1921 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     1981 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agttctagac
     2041 aaacaccata caacaagaaa caaacaccat acaacaagaa agggccctag agtcggggcg
     2101 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2161 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2221 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2281 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2341 tcgataagga tctgaacgat ggagcggaga atgggcggaa ctgggcggag ttaggggcgg
     2401 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     2461 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     2521 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     2581 cacattccac agcggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2641 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2701 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2761 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2821 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3121 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3481 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4801 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5101 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5161 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5221 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5281 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     5341 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     5401 tatcgatagg taccgagctc ttacgcgtgc tagcccgggc tcgagatctg cgatc
//
