LOCUS       sequence-217328-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses worm ACP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  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 6748)
  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: ACP2-V5
FEATURES             Location/Qualifiers
     source          1..6748
                     /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    2315..2436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2326..2345)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2380..2399
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2622..3077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2709..2728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2919..2940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3194..3551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3194..3385
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3217..3237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3402..3537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3602..4396
                     /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(3656..3675)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4266..4285
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4460..4508
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4549..4568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4668..4690
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4728..4746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4814..4918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4919..4987,4988..5779)
                     /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(5137..5156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5950..6538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6439..6458
                     /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 attcacgcgt ggtacctcta gagtcgagcc accatgaaga tttcatggtt tttgttcatt
     1021 tgtttgattg ttcaattcgc catcgcactt tctgaaaacc ccaaacggat ttacacacaa
     1081 gtgctatttc gtcatggagc ccgtgcccca tcaaatagtc tcagcgatcc tgcttatctt
     1141 gcaaactttc caagaggtct aggagaattg actgatagag gcttcgaaaa ttcatttcga
     1201 ttgggacgat tcttaaaaac tcgatatgta gactcgaaat ttgttgatgg aaatttgaag
     1261 ccacgtcaga tgtattggag atccgtaaac aaaaatcgat gtctaagctc agcgtcaacc
     1321 gttggtgcgg ctatgttcga agatccatct aggcatttat atgttcctgt tcttacagag
     1381 gagatcggag aaaatctact aaactacgac caagctaact gtccccgaga gttagagctg
     1441 atcaaagaga aatgcccaga ttttggtgga agttttcatc cgtggacgat ttacgaggaa
     1501 ttcattgcca actgtcttaa atatactcac ccagttttca agcaataccc ttttcacacc
     1561 attgaagctc atatcaatga gtacaaaaat ggaattcctc ttccaccact catcgctcaa
     1621 catataaacg aaatcatggg tatttatgtg aatgtgaccc agtttatcac aggaactggt
     1681 aatcatcatg acccgcgaat gatgaaggtc aaatttggaa atcttatgaa cacgttgcta
     1741 acggacatta aaaataagat atacatggat aaagaagaga aacacaagag tgatggaaga
     1801 gtatccagaa gggagaagtt ggcagtgtac tctacgcaag actggattct aatgggagtg
     1861 ctcgattcat tgaatgtgtt ggaacaaact gttggactcg acaaatatcc cgaatacaac
     1921 tcaatgatca tatttgaaac gtggaaggat aatggaaaat attttgtgaa agttttctat
     1981 aaaaaagaag agatcactgc cgaggatcat gagctaatag atgtcaccaa atttgtgaga
     2041 aactgtaaaa gagaacgatg tttggcacag gatttcctcg actgctgcga tgattatcga
     2101 aacaaaggtg aagctgagag ttgtgaagca tctgatttaa agattcgaac cacgagaagc
     2161 tctccaccta ataatattag aagagatctt catgtatcgg aattctctgg tccagaagtc
     2221 gacggcaaac caatcccaaa cccactgctg ggcctggatt aaggatccgc ggccgcttcc
     2281 ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa
     2341 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2401 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2461 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2521 tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg
     2581 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     2641 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2701 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2761 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2821 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2881 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2941 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3001 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3061 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3121 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     3181 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3481 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3541 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac
     3601 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3661 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3721 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3781 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3841 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3901 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3961 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4021 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4081 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4141 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4201 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4261 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4321 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4381 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4441 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     4501 tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc
     4561 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4621 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4681 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4741 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4801 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4921 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4981 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5041 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5101 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5161 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5221 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5281 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5341 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5401 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5461 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5521 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5581 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5641 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5701 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5761 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5821 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5881 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5941 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6001 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6061 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc
     6601 tcgacagatc ttcaatattg gccattagcc atattattca ttggttatat agcataaatc
     6661 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6721 ggctcatgtc caatatgacc gccatgtt
//
