LOCUS       sequence-190581-        5238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ACP-Tagged GPI anchor in Mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5238)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5238)
  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            "Unpublished"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: pACP-GPI Control Plasmid
FEATURES             Location/Qualifiers
     source          1..5238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             765..998
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     primer_bind     complement(1145..1164)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1146..1164)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1438..1559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1475..1494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1529..1548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      1587..1614
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1633..1724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1725..1793,1794..2585)
                     /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     complement(1943..1962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        2748..3105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2748..2939
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2771..2791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2956..3091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3140..3934
                     /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             3140..3934
                     /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(3194..3213)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3804..3823
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4122..4141)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4166..4213
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4542..5130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5031..5050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgcgatatc accatgggaa tccaaggagg gtctgtcctg ttcgggctgc
      721 tgctcgtcct ggctgtcttc tgccattcag gtcatagcag atctatgagc actatcgaag
      781 aacgcgttaa gaaaattatc ggcgaacagc tgggcgttaa gcaggaagaa gttaccaaca
      841 atgcttcttt cgttgaagac ctgggcgcgg attctcttga caccgttgag ctggtaatgg
      901 ctctggaaga agagtttgat actgagattc cggacgaaga agctgagaaa atcaccaccg
      961 ttcaggctgc cattgattac atcaacggcc accaggcgga attccttgaa aatggtggga
     1021 catccttatc agagaaaaca gttcttctgc tggtgactcc atttctggca gcagcctgga
     1081 gccttcatcc ctaaggatcc tgactcgagg gggggcccgg taccaggtaa gtgtacccaa
     1141 ttcgccctat agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag
     1201 cctgaatggc gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta
     1261 attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac
     1321 agtctgttca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     1381 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     1441 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1501 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     1561 cgcgtaaatt gtaagcgtta ataagcagaa ggccatcctg acggatggtc tttttgcgtt
     1621 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2641 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2701 gtgagttttc gttccactga gcgtcagaca ttgaaaaagg aagaatcctg aggcggaaag
     2761 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3121 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3181 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3241 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3301 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3361 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3421 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3481 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3541 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3601 tcaggatgat ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3661 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3721 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3781 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg
     3841 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3901 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3961 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4021 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4081 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4141 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4201 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4261 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4321 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4381 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5221 gataaccgta ttaccgcc
//
