LOCUS       sequence-217206-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human prostatic acid phosphatase (isoform 1) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  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 6648)
  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: ACPPiso1-V5
FEATURES             Location/Qualifiers
     source          1..6648
                     /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    2215..2336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2226..2245)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2280..2299
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2522..2977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2609..2628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2819..2840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3094..3451
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3094..3285
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3117..3137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3302..3437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3502..4296
                     /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(3556..3575)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4166..4185
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4360..4408
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4449..4468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4568..4590
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4628..4646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4819..4887,4888..5679)
                     /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(5037..5056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5850..6438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6339..6358
                     /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 catcaaggta
     2101 ctgaagacag tacagatgtc gacggcaaac caatcccaaa cccactgctg ggcctggatt
     2161 aaggatccgc ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat
     2221 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2281 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2341 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     2401 gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa
     2461 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2521 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2941 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3001 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3061 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3361 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3421 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     3481 gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc
     3541 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3601 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3661 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3721 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3781 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3841 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3901 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3961 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4021 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4081 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4141 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4201 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4261 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4321 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     4381 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc
     4441 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4501 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4561 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4621 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4681 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5041 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6481 tggccttttg ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca
     6541 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     6601 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
