LOCUS       sequence-142854-        7356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SLC24A4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  AUTHORS   Li XF, Kraev AS, Lytton J
  TITLE     Molecular cloning of a fourth member of the potassium-dependent 
            sodium-calcium exchanger gene family, NCKX4.
  JOURNAL   J Biol Chem. 2002 Dec 13;277(50):48410-7. Epub 2002 Oct 11.
  PUBMED    12379639
REFERENCE   2  (bases 1 to 7356)
  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: "J Biol 
            Chem."; date: "2002-12-13"; volume: "277(50)"; pages: "48410-7. Epub
            2002 Oct 11"
COMMENT     Sequence Label: Mouse NCKX4-AS-long (pcDNA3.1+)
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2946..2963)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2952..3176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3222..3650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3309..3328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3519..3540
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3659..3679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3664..3994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3845..3980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3907..3926
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4061..4855
                     /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(4115..4134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4725..4744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5029..5150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5066..5085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5120..5139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5199..5215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5199..5215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5212..5234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5223..5239
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5247..5253,5254..5271,5272..5277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5292..5313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5430..5447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5601..6189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5681..5700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6360..7151,7152..7220))
                     /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     6983..7002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7221..7325)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccttgaagct cgggatcttc ttccaggggc gcggggcgta
      961 cggagatggc tctccgtgga ctcatccggc agagcaaagt tcgccggagg cgagagatgc
     1021 tgccgcagca agttggcttc gtgtgcgctg tgctggccct ggtgtgctgt gcctccggcc
     1081 tctttggaag cttgggccac aaaacagctt ctgctggcaa acatgttctg ctggacacgt
     1141 ggcgaaatag aaaattgatg gcaccaatca atgggactcc gctggctaag aactgcacag
     1201 atcctgcgat tcacgagttc cccacagacc tgttctccaa caaagaacga cagcacggcg
     1261 ctgtcctgct acacatcctt ggtgctctct atatgttcta tgccctggcc atcgtgtgcg
     1321 atgacttctt tgttccatcc ctagagaaga tctgtgagaa acttcatctg agtgaagatg
     1381 tggcaggtgc caccttcatg gctgcaggaa gctcgacacc agagctgttt gcttctgtca
     1441 ttggtgtgtt catcacccac ggagatgtcg gtgtaggaac gattgtgggc tctgctgtgt
     1501 tcaacatcct atgtatcatt ggagtgtgtg gactctttgc tggacaggtg gtgcggttga
     1561 cgtggtgggc tgtgtgccgt gactctgtgt actacacgct ctctgtcatt gtgctcattg
     1621 cgttcatata cgatgaggaa attgtgtggt gggaaggcct ggtgctcatc atcctgtacg
     1681 tgttttatat tcttatcatg aagtacaata tgaagatgca gacatttttc acaaccaaac
     1741 aaaagagcat tgcaaatggt aacccggtca gcaacgagct ggaagatggt aatgacctct
     1801 atgatggtag ctatgacgac ccttctgtgc cattgctggg gcaagtgaag gagaaacctc
     1861 cctatggcaa aacccccgtg gttatggtgg atgagattct cagctctagc cctcccaagt
     1921 tcaccttccc tgaggctggc ctgcgcatca tgatcaccaa caagtttggg cccaggactc
     1981 gactacggat ggccagcagg atcataatta atgaacgaca gaggttgatc aactcagcca
     2041 atggtgttaa cagcaagcca ctgcaaaacg ggagacatga gaacatggag aatggaaatg
     2101 ttcctgtgga gaaccctgaa gacccacagc aaggccagga gcagcagccg ccaccacagc
     2161 caccaccccc agagccagag tcagtagaga cagtcttcct gtcacccttc tcaatgcctg
     2221 aggcaaaagg cgacaaggct aagtgggtgt tcacgtggcc cctcatcttc ctcctgtgtg
     2281 tcaccatccc caactgcagc aagccccgct gggagaagtt cttcatggtc accttcatca
     2341 cggccactct gtggattgcc gtgttctcct acctcatggt gtggctggtg actattattg
     2401 gatacacact tgggatcccc gatgtcatca tgggcatcac cttcctggcg gcaggcacaa
     2461 gtgttccaga ctgcatggca agcttaattg tggcaagaca aggtcttgga gacatggcag
     2521 tctctaacac catcggaagc aacgtgtttg atatcctggt ggggcttggc atcccgtggg
     2581 gcctgcagac catggttatt aattatggat ccacagtgaa gatcaacagc cggggtctgg
     2641 tctattccgt ggtgctgctg ctgggctctg tcgctctcac tgtccttggt attcacctca
     2701 acaaatggcg cctggaccgg aaactgggca tctacgtgct ggtcctctat gctgtcttcc
     2761 tgtgcttctc cataatgata gagtttaatg tctttacctt cgtcaacttg cccatgtgca
     2821 gagaagacga ctaaagctga accattgccc aggaaactgc tgttgccacc gagatgaatt
     2881 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
     2941 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3001 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3061 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3121 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3181 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3241 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3301 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3361 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3421 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3481 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3541 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3601 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3901 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3961 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4021 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4081 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4141 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4201 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4261 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4321 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4381 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4441 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4501 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4561 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4621 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4681 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4741 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4801 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4861 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4921 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4981 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5041 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5101 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5161 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5221 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5281 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5341 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5401 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5461 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5521 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5581 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5641 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5701 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5761 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5821 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5881 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5941 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6001 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6061 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6121 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6181 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6241 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6301 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6361 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6421 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6481 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6541 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6601 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6661 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6721 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6781 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6841 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6901 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6961 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7021 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7081 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7141 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7201 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7261 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7321 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
