LOCUS       sequence-80505-e        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a bicistronic IRES-mCherry containing vector for mammalian 
            expression (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  TITLE     Vignali lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6754)
  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            "Unpublished"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: pICherryNeo
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1151..1699
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1295..1312)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1522..1541
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1702..2409
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAG
                     GRHSTGGMDELYK"
     primer_bind     complement(1844..1862)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2112..2131
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2457..2578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2468..2487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2522..2541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2764..3219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2851..2870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3061..3082
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3336..3693
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3336..3527
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3359..3379)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3544..3679
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3744..4538
                     /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(3798..3817)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4408..4427
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4602..4650
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4692..4711)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4811..4833
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4871..4889)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4957..5061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5062..5130,5131..5922)
                     /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(5280..5299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6093..6681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6582..6601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt cacgcgtggt acctctagag tcgagcggga tcaattccgc
     1141 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1201 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1261 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1321 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1381 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1441 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1501 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     1561 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     1621 tgtgtttagt cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     1681 tttgaaaaac acgataatac catggtgagc aagggcgagg ataacatggc catcatcaag
     1741 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1801 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1861 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1921 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     1981 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     2041 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     2101 ccctccgacg gccccgtaat gcagaagaag acgatgggct gggaggcctc ctccgagcgg
     2161 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     2221 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     2281 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     2341 atcgtggaac agtacgaacg cgccgggggc cgccactcca ccggcggcat ggacgagctg
     2401 tacaagtaaa gcggccgctt ccctttagtg agggttaatg cttcgagcag acatgataag
     2461 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2521 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2581 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     2641 aagcaagtaa aacctctaca aatgtggtaa aatccgataa ggatcgatcc gggctggcgt
     2701 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2761 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2821 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2881 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2941 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3001 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3061 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3121 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3181 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     3241 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     3301 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     3361 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3481 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3541 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3601 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3661 ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc
     3721 tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg
     3781 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3841 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3901 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3961 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4021 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4081 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4141 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4201 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4261 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4321 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4381 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4441 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4501 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4561 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     4621 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc gcgcgtatgg
     4681 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4741 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4801 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4861 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4921 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5281 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5581 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6721 tgctggcctt ttgctcacat ggctcgacag atct
//
