LOCUS       sequence-194153-        5088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid that allows for the expression of a 
            gene of interest fused to the N-terminus or C-terminus of MCP-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5088)
  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            "Unpublished"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: pMCP-tag (m) Vector
FEATURES             Location/Qualifiers
     source          1..5088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..89
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(275..1066,1067..1135))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     898..917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1136..1227)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1246..1273)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     polyA_signal    1301..1422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1312..1331)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1366..1385
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1696..1715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1696..1714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1787..2020)
                     /codon_start=1
                     /product="mutant acyl carrier protein (ACP) from E. coli"
                     /label=MCP
                     /note="The mutations (D36T and D39G) prevent recognition by
                     ACP synthase while allowing recognition by SFP synthase."
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGATSLGTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     promoter        complement(2072..2090)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2136..2339)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2165..2185)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2340..2643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2818..3406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2898..2917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3735..3782
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3807..3826
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4014..4808)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4014..4808)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4125..4144)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4735..4754
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4843..5088,1..112))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4857..4992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(5009..5088,1..112)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
       61 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggattcttc
      121 ctttttcaat gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
      901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1201 ttgaatgtat ttagaaaaat aaacaaaaga gtttgtagaa acgcaaaaag accatccgtc
     1261 aggatggcct tctgcttatt aacgcttaca atttacgcgt taagatacat tgatgagttt
     1321 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1381 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1441 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1501 tacaaatgtg gtatggctga ttatgatcat gaacagactg tgaggactga ggggcctgaa
     1561 atgagccttg ggactgtgaa tctaaaatac acaaacaatt agaatcagta gtttaacaca
     1621 ttatacactt aaaaattgga tctccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1681 cgatcgagtg aattgtaata cgactcacta tagggcgaat tgggtacact tacctggtac
     1741 cgggcccccc ctcgagtcag gatcctggcg cgcctatacc tgcaggcgcc tggtggccgt
     1801 tgatgtaatc aatggcagcc tgaacggtgg tgattttctc agcttcttcg tccggaatct
     1861 cagtatcaaa ctcttcttcc agagccatta ccagctcaac ggtcccaaga gaagtcgcgc
     1921 ccaggtcttc aacgaaagaa gcattgttgg taacttcttc ctgcttaacg cccagctgtt
     1981 cgccgataat tttcttaacg cgttcttcga tagtgctcat ggtaagcttg atatcgcggc
     2041 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     2101 cgtaatcgct agcggatctg acggttcact aaaccagctc tgcttatata gacctcccac
     2161 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     2221 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     2281 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     2341 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     2401 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     2461 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     2521 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     2581 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     2641 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     2701 ctaatgcatg gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3541 aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc
     3601 tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg
     3661 ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     3721 aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc
     3781 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct
     3841 agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     3901 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     3961 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
     4021 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     4081 agcacgagga agcggtcagc ccattcgccg ccaagttctt cagcaatatc acgggtagcc
     4141 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     4201 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     4261 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     4321 tgatgttctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     4381 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     4441 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     4501 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     4561 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     4621 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     4681 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     4741 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     4801 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc
     4861 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     4921 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4981 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     5041 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacct
//
