LOCUS       sequence-142856-        7406 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 7406)
  TITLE     Lytton lab plasmids
REFERENCE   2  (bases 1 to 7406)
  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"
COMMENT     Sequence Label: Rat NCKX4-AS-short (pcDNA3.1+)
FEATURES             Location/Qualifiers
     source          1..7406
                     /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"
     CDS             2903..2944
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2954..2971
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2994..3011)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3000..3224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3270..3698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3357..3376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3567..3588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3707..3727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3712..4042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3893..4028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3955..3974
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4109..4903
                     /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(4163..4182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4773..4792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5079..5200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5170..5189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5249..5265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5249..5265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5262..5284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5273..5289
                     /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(5297..5303,5304..5321,5322..5327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5342..5363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5480..5497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5651..6239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5731..5750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6410..7201,7202..7270))
                     /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     7033..7052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7271..7375)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttccttgaa
      961 gctccggatc ttcttccagg gacaccgggc agacggagat ggcgctccgt ggactcatcc
     1021 ggcattgcaa agttcgccgg aggcgagaga tgctgccgca gcaagttggc ttcgtgtgcg
     1081 ctgtgttcgc cctggtgtgc tgtgcctccg gcctctttgg aagcttgggc cacaaaacag
     1141 cttctgctgg caaacacgtt ctgccagaca cgtggcgaaa tagaaagttg atggcaccga
     1201 taaatgggac tccactggct aagaactgta cagatcctgc gatccacgag ttccctacag
     1261 acctgttctc caacaaagaa cgacagcatg gcgctgtcct gctacacatc cttggtgcac
     1321 tctatatgtt ctacgccctg gccatcgtgt gtgatgactt ctttgtccca tccctagaga
     1381 agatctgtga gaaacttcat ctgagtgaag atgtggcggg tgccaccttc atggctgcag
     1441 gaagctcgac accagagctg tttgcttctg tcattggtgt gttcatcacc catggagatg
     1501 ttggtgtagg aacgattgtg ggctcagctg tgttcaacat cctatgtatc attggagtgt
     1561 gtggactctt tgctggacag gtggtgcggt tgacgtggtg ggctgtgtgc cgtgactctg
     1621 tgtactacac actctctgtc atcgttctca ttgcgttcat atacgatgag gaaattgtgt
     1681 ggtgggaagg tctgatcctc atcatcttgt acgtgtttta tattctcatc atgaagtaca
     1741 atatgaagat gcagacgttt ttcacaacca aacaaaagag cattgcaaat ggtaacccag
     1801 tcagcaacga gctggaagat gtgaaggaga aacctcccta tggcaagtcc cccgtggtga
     1861 tggtggatga gattctcagc tccagtcctc ccaagttcac cttccctgag gctggcctgc
     1921 gcatcatgat caccaacaag tttgggccca ggacccgact acggatggcc agcaggatca
     1981 taattaatga acgacagagg ttgatcaact cggccaatgg tgttaacagc aagccactgc
     2041 aaaatgggag acacgagaac atggagaatg gaaatgtccc tgtagagaac cctgaagacc
     2101 cacagcaggg ccaggagcag cagccgccac cacagccgcc acccccagag cctgagtcgg
     2161 tagagacggt cttcctgtcg cccttctcaa tgcctgaggc aaaaggcgac aaggctaagt
     2221 gggtgttcac ttggcccctc atcttcctcc tgtgcgtcac catccccaac tgcagcaagc
     2281 ctcgctggga gaagttcttc atggtcacct tcatcacggc cactctgtgg atcgcggtgt
     2341 tctcctacct catggtgtgg ctggtgacta ttattggata cacacttggg atccctgatg
     2401 tcatcatggg catcactttc ctggcggcag gcacgagtgt cccagactgc atggccagct
     2461 taattgtggc gagacaaggg cttggcgaca tggcagtctc taacaccatc ggaagcaacg
     2521 tgtttgacat cctggtgggg cttggcatcc catggggcct gcagaccatg gttattaatt
     2581 atggatccac agtgaagatc aacagccggg ggctggtcta ttccgtggtg ctgctgctgg
     2641 gctctgtggc tctcactgtc cttggcatcc acctcaacaa atggcgcctg gacaggaagc
     2701 tgggcatcta cgtgctggtc ctctatgccg tctttctgtg cttctcaata atgatagagt
     2761 ttaacgtctt taccttcgtc aacttgccca tgtgcagaga agacgactaa agctgagctg
     2821 tcacccagga aagtgcaagg gcaattctgc agatatccag cacagtggcg gccgctcgag
     2881 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     2941 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     3001 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3061 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3121 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3181 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3241 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3301 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3361 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3421 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3481 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3541 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3601 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3661 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3721 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
     3781 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3841 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3901 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3961 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4021 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4081 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4141 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4201 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4261 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4321 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4381 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4441 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4501 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4561 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4621 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4681 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4741 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4801 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4861 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4921 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5041 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5221 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5281 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5341 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5401 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5461 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5821 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6061 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6181 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6241 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6301 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6361 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6421 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6481 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6541 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6601 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6661 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6721 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6781 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6841 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6901 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6961 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7021 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7081 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7141 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7201 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7261 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7321 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7381 ttccccgaaa agtgccacct gacgtc
//
