LOCUS       sequence-142860-        7299 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 7299)
  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 7299)
  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: Human NCKX4-AS-short-FlagTag (pcDNA3.1+)
FEATURES             Location/Qualifiers
     source          1..7299
                     /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     934..950
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1109..1132
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2842..2858)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2889..2906)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2895..3119
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3165..3593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3252..3271)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3462..3483
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3602..3622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3607..3937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3788..3923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3850..3869
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4004..4798
                     /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(4058..4077)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4668..4687
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4972..5093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5009..5028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5063..5082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5142..5158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5142..5158)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5155..5177)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5166..5182
                     /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(5190..5196,5197..5214,5215..5220))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5235..5256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5373..5390)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5544..6132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5624..5643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6303..7094,7095..7163))
                     /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     6926..6945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7164..7268)
                     /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 ccgggccccc cctcgaggtc gacggtatcg ataagcttga
      961 tctcaaagtt cgcaggaggc gagagatgct gccgcagcaa gtcggcttcg tgtgcgcggt
     1021 gctggccctg gtgtgctgtg cgtccggcct cttcggcagc ttggggcaca aaacagcttc
     1081 tgctagcaaa cgtgtcctgc cagacacaga ctacaaggac gacgatgaca agtggagaaa
     1141 tagaaagttg atggccccag tgaatgggac acagacagcc aagaactgca cagatcctgc
     1201 gattcacgag ttccccacag atctgttctc caataaggag cgacagcacg gagccgtcct
     1261 gctgcacatc cttggtgctc tgtatatgtt ctatgccttg gccatagtgt gcgatgactt
     1321 ctttgttccg tctctagaga agatctgtga gagactccat ctgagcgaag atgtggctgg
     1381 agccaccttc atggctgcag gaagctcaac gccagagctg tttgcgtctg ttattggggt
     1441 gttcatcacc catggggacg tcggggtggg caccatcgtg ggctctgctg tgttcaacat
     1501 cctgtgcata attggagtgt gcggactgtt tgctggccag gtggtccgtc tgacgtggtg
     1561 ggccgtgtgc cgagactccg tgtactacac catctctgtc atcgtgctca tcgtgttcat
     1621 atatgatgaa caaattgtgt ggtgggaagg cctggtgctc atcatcttgt atgtgtttta
     1681 tattctgatc atgaagtaca atgtgaagat gcaagccttt ttcacagtca aacaaaagag
     1741 cattgcaaac ggtaacccgg tcaacagtga gctggaggct gtgaaggaga agccacagta
     1801 tggcaagaac cccgtggtga tggtggacga gattatgagc tccagccctc ccaagttcac
     1861 cttccctgaa gcaggcttac gaatcatgat caccaataag tttggaccta ggacccgact
     1921 acggatggcc agcaggatca tcattaatga gcggcagaga ctgatcaact cggccaatgg
     1981 tgtgagcagt aagccgcttc aaaacgggag gcacgagaac attgagaacg ggaatgttcc
     2041 tgtggaaaac cccgaagacc ctcagcagaa tcaggagcag cagccgccgc cacagccacc
     2101 accgccagag ccagagccgg tggaggctga cttcctgtcc cccttctccg tgccggaggc
     2161 cagaggggac aaggtcaagt gggtgttcac ctggcccctc atcttcctcc tgtgcgtcac
     2221 cattcccaac tgcagcaagc cccgctggga gaagttcttc atggtcacct tcatcaccgc
     2281 cacgctgtgg atcgctgtgt tctcctacat catggtgtgg ctggtgacta ttatcggata
     2341 cacacttggg atcccggatg tcatcatggg cattactttc ctggcagcag ggacaagtgt
     2401 tccagactgc atggccagcc taattgtggc gagacaaggc cttggggaca tggcagtctc
     2461 caacaccata ggaagcaacg tgtttgacat cctggtagga cttggtgtac cgtggggcct
     2521 gcagaccatg gttgttaatt atggatcaac agtgaagatc aacagccggg ggctggtcta
     2581 ttccgtggtc ctgttgctgg gctctgtcgc tctcaccgtc ctcggcatcc acctaaacaa
     2641 gtggcgactg gaccggaagc tgggtgtcta cgtgctggtt ctctacgcca tcttcttgtg
     2701 cttctccata atgatagagt ttaacgtctt taccttcgtc aacttgccga tgtgccggga
     2761 agacgattag cgctgagtcg cggcccctgg gagctgatct ggacaccctg tgacactggc
     2821 gttctcctct atcaagctta tcgataccgt cgacctcgag ggggggcccg tttaaacccg
     2881 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2941 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3001 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3061 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3121 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3181 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3241 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3301 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3361 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3421 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3481 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3541 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3601 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat
     3661 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3781 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3841 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3901 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3961 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4021 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4081 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4141 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4201 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4261 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4321 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4381 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4441 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4501 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4561 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4621 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4681 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4741 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4801 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4861 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5101 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5161 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5221 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5281 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5341 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5401 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5461 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5521 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5581 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5641 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5701 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5761 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5881 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5941 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6001 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6061 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6121 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6181 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6241 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6301 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6361 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6421 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6481 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6721 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6961 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7141 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7201 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7261 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
