LOCUS       sequence-194079-        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  JOURNAL   Unpublished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            "Unpublished"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        100..291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(454..1245,1246..1314))
                     /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     1077..1096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1315..1406)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1425..1452)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     polyA_signal    1480..1601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1491..1510)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1545..1564
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1875..1894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1875..1893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2041..2274)
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     promoter        complement(2401..2419)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2465..2668)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2494..2514)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2669..2972
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3147..3735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3227..3246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4064..4111
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4136..4155
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4343..5137)
                     /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(4343..5137)
                     /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(4454..4473)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5064..5083
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5172..5238,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5186..5238,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggattcttcc
      301 tttttcaatg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     1081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1381 tgaatgtatt tagaaaaata aacaaaagag tttgtagaaa cgcaaaaaga ccatccgtca
     1441 ggatggcctt ctgcttatta acgcttacaa tttacgcgtt aagatacatt gatgagtttg
     1501 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1561 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1621 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1681 acaaatgtgg tatggctgat tatgatcatg aacagactgt gaggactgag gggcctgaaa
     1741 tgagccttgg gactgtgaat ctaaaataca caaacaatta gaatcagtag tttaacacat
     1801 tatacactta aaaattggat ctccattcgc cattcaggct gcgcaactgt tgggaagggc
     1861 gatcgagtga attgtaatac gactcactat agggcgaatt gggtacactt acctggtacc
     1921 gggccccccc tcgagtcagg atccttaggg atgaaggctc caggctgctg ccagaaatgg
     1981 agtcaccagc agaagaactg ttttctctga taaggatgtc ccaccatttt caaggaattc
     2041 cgcctggtgg ccgttgatgt aatcaatggc agcctgaacg gtggtgattt tctcagcttc
     2101 ttcgtccgga atctcagtat caaactcttc ttccagagcc attaccagct caacggtgtc
     2161 aagagaatcc gcgcccaggt cttcaacgaa agaagcattg ttggtaactt cttcctgctt
     2221 aacgcccagc tgttcgccga taattttctt aacgcgttct tcgatagtgc tcatagatct
     2281 gctatgacct gaatggcaga agacagccag gacgagcagc agcccgaaca ggacagaccc
     2341 tccttggatt cccatggtga tatcgcggcc gccaccgcgg tggagctcca gcttttgttc
     2401 cctttagtga gggttaattt cgagcttggc gtaatcgcta gcggatctga cggttcacta
     2461 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     2521 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     2581 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     2641 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     2701 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     2761 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     2821 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     2881 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     2941 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     3001 atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg gttatccaca
     3061 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3121 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3181 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3241 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3301 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3361 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3421 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3481 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3541 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3601 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3661 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3721 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3781 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3841 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     3901 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     3961 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     4021 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     4081 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     4141 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     4201 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     4261 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     4321 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4381 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4441 caagttcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4501 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4561 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4621 tggcgaacag ttcggctggc gcgagcccct gatgttcttc gtccagatca tcctgatcga
     4681 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4741 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4801 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4861 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4921 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4981 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5041 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5101 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5161 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     5221 agctcagagg ccgaggcg
//
