LOCUS       Exported                7178 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of SLC8B1.
ACCESSION   .
VERSION     .
KEYWORDS    Human NCKX6 (pcDNA3.1+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7178)
  AUTHORS   Cai X, Lytton J
  TITLE     Molecular cloning of a sixth member of the K+-dependent Na+/Ca2+ 
            exchanger gene family, NCKX6.
  JOURNAL   J Biol Chem. 2004 Feb 13;279(7):5867-76. Epub 2003 Nov 18.
  PUBMED    14625281
REFERENCE   2  (bases 1 to 7178)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7178
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     protein_bind    1519..1532
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     primer_bind     complement(2726..2742)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2726..2742)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2768..2785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2774..2998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3044..3472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3131..3150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3341..3362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3481..3501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3486..3816
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3667..3802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3729..3748
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3883..4677
                     /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(3937..3956)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4547..4566
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4851..4972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4888..4907)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4942..4961
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5021..5037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5021..5037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5034..5056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5045..5061
                     /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(5069..5099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5114..5135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5252..5269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5423..6011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5503..5522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6182..7042)
                     /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     6805..6824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7043..7147)
                     /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 aagcttgata tggccggcag aaggctgaat ctgcgctggg cactgagtgt
      961 gctttgtgtg ctgctaatgg cggagacagt gtctgggact aggggctcgt ctacaggagc
     1021 tcacattagc ccccagtttc cagcttcagg tgtgaaccag acccccgtgg tagactgccg
     1081 caaggtgtgt ggcctgaatg tctctgaccg ctgtgacttc atccggacca accctgactg
     1141 ccacagtgat ggggggtacc tggactacct ggaaggcatc ttctgccact tccctcccag
     1201 cctcctccct ctggctgtca ctctctacgt ttcctggctg ctctacctgt ttctgattct
     1261 gggagtcacc gcagccaagt ttttctgccc caacttgtcg gccatttcta ccacactgaa
     1321 gctctcccac aacgtggcag gcgtcacctt cctggcattt gggaatggtg cacctgacat
     1381 cttcagtgcc ctggtggcct tctctgaccc gcacacagcc ggcctggccc ttggggcact
     1441 gtttggcgct ggcgtgctgg ttaccacagt ggtggccgga ggcattacca tcctacaccc
     1501 cttcatggct gcctccaggc ccttcttcag ggacatcgtt ttctacatgg tggctgtgtt
     1561 cctgaccttc ctcatgctct tccgtggcag ggtcaccctg gcatgggctc tgggttacct
     1621 gggcttgtat gtgttctatg tggtcactgt gattctctgc acctggatct accaacggca
     1681 acggagagga tctctgttct gccccatgcc agttactcca gagatcctct cagactccga
     1741 ggaggaccgg gtatcttcta ataccaacag ctatgactac ggtgatgagt accggccgct
     1801 gttcttctac caggagacca cggctcagat cctggtccgg gccctcaatc ccctggatta
     1861 catgaagtgg agaaggaaat cagcatactg gaaagccctc aaggtgttca agctgcctgt
     1921 ggagttcctg ctgctcctca cagtccccgt cgtggacccg gacaaggatg accagaactg
     1981 gaaacggccc ctcaactgtc tgcatctggt tatcagcccc ctggttgtgg tcctgaccct
     2041 gcagtcgggg acctatggtg tctatgagat aggcggcctc gttcccgtct gggtcgtggt
     2101 ggtgatcgca ggcacagcct tggcttcagt gacctttttt gccacatctg acagccagcc
     2161 ccccaggctt cactggctct ttgctttcct gggctttctg accagcgccc tgtggatcaa
     2221 cgcggccgcc acagaggtgg tgaacatctt gcggtccctg ggtgtggtct tccggctgag
     2281 caacactgtg ctggggctca cgctgctggc ctgggggaac agcattggag atgccttctc
     2341 ggatttcaca ctggctcgcc agggctaccc acggatggcg ttctccgcct gctttggcgg
     2401 catcatcttc aacatcctcg tgggtgtggg gctgggctgc ctgctccaga tctcccgaag
     2461 ccacacagaa gtgaagctgg agccagacgg actgctggtg tgggtcctgg caggcgccct
     2521 ggggctcagc ctcgtcttct ccctggtctc agtcccattg cagtgcttcc agctcagcag
     2581 agtctatggc ttctgcctgc tcctcttcta cctgaacttc cttgtcgtgg ccctcctcac
     2641 tgaatttgga gtgattcacc tgaaaagcat gtgactgaag ccgcttagtg ctgtggcctc
     2701 actatcgaat tcctgcagcc cgggggatcc actagttcta gagggcccgt ttaaacccgc
     2761 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2821 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2881 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2941 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3001 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3061 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3121 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3181 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3241 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3301 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3361 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3421 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg
     3541 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3601 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3661 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3721 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3781 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3841 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3901 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3961 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4021 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4081 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4141 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4201 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4261 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4321 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4381 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4441 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4501 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4561 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4621 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4681 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4741 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4801 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4861 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4921 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4981 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5041 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5101 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5161 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5221 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5281 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5341 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5401 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5461 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5521 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5581 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5641 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5701 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5761 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5821 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5881 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5941 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6001 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6061 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6121 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6181 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6241 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6301 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6361 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6421 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6481 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6541 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6601 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6661 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6721 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6781 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6841 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6901 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6961 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7021 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7081 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7141 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
