LOCUS       sequence-217204-        6744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human prostatic acid phosphatase (isoform 2) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   Cotella D, Hernandez-Enriquez B, Duan Z, Wu X, Gazula VR, Brown MR, 
            Kaczmarek LK, Sesti F
  TITLE     An evolutionarily conserved mode of modulation of Shaw-like K(+) 
            channels.
  JOURNAL   FASEB J. 2013 Apr;27(4):1381-93. doi: 10.1096/fj.12-222778. Epub 
            2012 Dec 11.
  PUBMED    23233530
REFERENCE   2  (bases 1 to 6744)
  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"; doi: 
            "10.1096/fj.12-222778"; journalName: "FASEB J"; date: "2013-04"; 
            volume: "27"; issue: "4"; pages: "1381-93"
COMMENT     Sequence Label: ACPPiso2-V5
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2311..2432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2322..2341)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2376..2395
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2618..3073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2705..2724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2915..2936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3190..3547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3190..3381
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3213..3233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3398..3533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3598..4392
                     /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(3652..3671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4262..4281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4456..4504
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4545..4564)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4664..4686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4724..4742)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4810..4914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4915..4983,4984..5775)
                     /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(5133..5152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5946..6534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6435..6454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
      961 tgagagctgc acccctcctc ctggccaggg cagcaagcct tagccttggc ttcttgtttc
     1021 tgcttttttt ctggctagac cgaagtgtac tagccaagga gttgaagttt gtgactttgg
     1081 tgtttcggca tggagaccga agtcccattg acacctttcc cactgacccc ataaaggaat
     1141 cctcatggcc acaaggattt ggccaactca cccagctggg catggagcag cattatgaac
     1201 ttggagagta tataagaaag aggtatagaa aattcttgaa tgagtcctat aaacatgaac
     1261 aggtttatat tcgaagcaca gacgttgacc ggactttgat gagtgctatg acaaacctgg
     1321 cagccctgtt tcccccagaa ggtgtcagca tctggaatcc tatcctactc tggcagccca
     1381 tcccggtgca cacagttcct ctttctgaag atcagttgct atacctgcct ttcaggaact
     1441 gccctcgttt tcaagaactt gagagtgaga ctttgaaatc agaggaattc cagaagaggc
     1501 tgcaccctta taaggatttt atagctacct tgggaaaact ttcaggatta catggccagg
     1561 acctttttgg aatttggagt aaagtctacg accctttata ttgtgagagt gttcacaatt
     1621 tcactttacc ctcctgggcc actgaggaca ccatgactaa gttgagagaa ttgtcagaat
     1681 tgtccctcct gtccctctat ggaattcaca agcagaaaga gaaatctagg ctccaagggg
     1741 gtgtcctggt caatgaaatc ctcaatcaca tgaagagagc aactcagata ccaagctaca
     1801 aaaaacttat catgtattct gcgcatgaca ctactgtgag tggcctacag atggcgctag
     1861 atgtttacaa cggactcctt cctccctatg cttcttgcca cttgacggaa ttgtactttg
     1921 agaaggggga gtactttgtg gagatgtact atcggaatga gacgcagcac gagccgtatc
     1981 ccctcatgct acctggctgc agccccagct gtcctctgga gaggtttgct gagctggctg
     2041 gccctgtgat ccctcaagac tggtccacgg agtgtatgac cacaaacagc catcaagttc
     2101 taaaggtcat ctttgctgtt gccttttgcc tgatatctgc tgtcctaatg gtactactgt
     2161 ttatccacat tcgccgtgga ctctgctggc agagagaatc ctatgggaac atcgtcgacg
     2221 gcaaaccaat cccaaaccca ctgctgggcc tggattaagg atccgcggcc gcttcccttt
     2281 agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2341 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2401 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2461 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2521 gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc gcaccgatcg
     2581 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     2641 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2701 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2761 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2821 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2881 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2941 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3001 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3061 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3121 gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
     3181 aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3541 ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag agccaccatg
     3601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4441 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4501 tgtgaatcga tagcgataag gatccgcgta tggtgcactc tcagtacaat ctgctctgat
     4561 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4621 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4681 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4741 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4801 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4861 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4921 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4981 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5041 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5101 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5161 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5221 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5281 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5341 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5401 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5461 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5521 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5581 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5641 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5701 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5761 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5821 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5881 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6001 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     6601 cagatcttca atattggcca ttagccatat tattcattgg ttatatagca taaatcaata
     6661 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     6721 catgtccaat atgaccgcca tgtt
//
