LOCUS       sequence-260086-        6000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 6000)
  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"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221_SLC12A6
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4125..4224)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4241..4260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4242..4260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4265..4281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4265..4281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4383..4402)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4394..5203
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4461..4480)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatgcacc cccctgagac cacaaccaag atggccagcg tgagattcat
      721 ggtgacaccc accaagatcg acgatatccc tggcctgtcc gacacatctc cagatctgag
      781 ctccaggtct agctccaggg tgcgcttttc tagcagagag agcgtgcccg agacctccag
      841 gtctgagcct atgagcgaga tgtccggagc aacaacctcc ctggcaacag tggccctgga
      901 cccaccctct gatcgcacca gccaccccca ggacgtgatc gaggatctgt ctcagaacag
      961 catcacaggc gagcacagcc agctgctgga cgatggccac aagaaggccc ggaacgccta
     1021 cctgaacaat tccaattatg aggagggcga cgagtacttc gataagaacc tggccctgtt
     1081 tgaggaggag atggacaccc gccccaaggt gtcctctctg ctgaatcgga tggccaacta
     1141 tacaaatctg acccagggag caaaggagca cgaggaggca gagaacatca cagagggcaa
     1201 gaagaagccc acaaagaccc ctcagatggg caccttcatg ggcgtgtacc tgccttgcct
     1261 gcagaatatc ttcggcgtga tcctgtttct gcggctgaca tgggtggtgg gaaccgcagg
     1321 cgtgctgcag gcctttgcca tcgtgctgat ctgctgttgc tgtacaatgc tgaccgccat
     1381 cagcatgtcc gccatcgcaa ccaacggagt ggtgccagca ggaggcagct atttcatgat
     1441 ctccagagcc ctgggacctg agtttggagg agcagtgggc ctgtgcttct atctgggcac
     1501 aacctttgcc gccgccatgt acatcctggg cgccatcgag atcttcctgg tgtacatcgt
     1561 gccaagggcc gccatctttc actctgacga tgccctgaag gagagcgccg ccatgctgaa
     1621 caatatgaga gtgtatggca ccgccttcct ggtgctgatg gtgctggtgg tgtttatcgg
     1681 cgtgaggtac gtgaacaagt tcgcctctct gtttctggcc tgcgtgatcg tgagcatcct
     1741 ggccatctat gccggcgcca tcaagagctc cttcgcccct ccacactttc ccgtgtgcat
     1801 gctgggcaat cgcaccctgt ctagccggca catcgacgtg tgcagcaaga caaaggagat
     1861 caacaatatg accgtgccat ccaagctgtg gggcttcttt tgcaactcct ctcagttctt
     1921 taatgccaca tgtgatgagt acttcgtgca caacaatgtg acctccatcc agggaatccc
     1981 aggactggca tctggcatca tcaccgagaa cctgtggagc aattatctgc caaagggcga
     2041 gatcatcgag aagccatccg ccaagagctc cgacgtgctg ggctctctga accacgagta
     2101 cgtgctggtg gatatcacaa cctcctttac actgctggtg ggcatcttct ttccttctgt
     2161 gaccggcatc atggccggct ccaatcgctc tggcgacctg aaggatgccc agaagagcat
     2221 cccaatcggc acaatcctgg ccatcctgac aacctctttc gtgtacctga gcaacgtggt
     2281 gctgtttgga gcatgcatcg agggagtggt gctgagggac aagttcggcg atgccgtgaa
     2341 gggcaatctg gtggtgggca ccctgagctg gccttcccca tgggtcatcg tgatcggctc
     2401 tttctttagc acatgtggag caggactgca gagcctgacc ggagcaccta gactgctgca
     2461 ggcaatcgcc aaggacaaca tcatcccatt cctgagggtg tttggccact ccaaggccaa
     2521 tggcgagcca acatgggcac tgctgctgac cgcagcaatc gcagagctgg gcatcctgat
     2581 cgccagcctg gatctggtgg cccctatcct gtccatgttc tttctgatgt gctatctgtt
     2641 cgtgaacctg gcctgtgccc tgcagacact gctgcgcacc ccaaattggc ggcccagatt
     2701 caggtactat cactgggccc tgtcctttat gggcatgtct atctgcctgg ccctgatgtt
     2761 tatctctagc tggtactatg ccatcgtggc catggtcatc gccggcatga tctacaagta
     2821 tatcgagtac cagggagcag agaaggagtg gggcgacggc atccggggcc tgtctctgag
     2881 cgccgcaaga ttcgcactgc tgaggctgga ggagggacca cctcacacca agaactggag
     2941 gccacagctg ctggtgctgc tgaagctgga cgaggatctg cacgtgaagc acccccggct
     3001 gctgacattc gcctctcagc tgaaggcagg caagggactg accatcgtgg gaagcgtgat
     3061 cgtgggcaac tttctggaga attacggaga ggccctggca gcagagcaga caatcaagca
     3121 cctgatggag gccgagaagg tgaagggctt ctgccagctg gtggtggccg ccaagctgag
     3181 agagggcatc agccacctga tccagtcctg tggcctgggc ggcatgaagc acaacaccgt
     3241 ggtcatgggc tggcctaatg gatggagaca gagcgaggac gcaagggcat ggaagacatt
     3301 catcggaacc gtgagggtga caaccgcagc acacctggcc ctgctggtgg caaagaacat
     3361 ctccttcttt ccatctaatg tggagcagtt ttccgagggc aacatcgacg tgtggtggat
     3421 cgtgcacgat ggcggcatgc tgatgctgct gcccttcctg ctgaagcagc acaaagtgtg
     3481 gcgcaagtgt tccatccgga tctttacagt ggcccagctg gaggacaatt ctatccagat
     3541 gaagaaggat ctggccacct tcctgtacca cctgagaatc gaggccgagg tggaggtggt
     3601 ggagatgcac gactccgata tctctgccta cacatatgag cgcaccctga tgatggagca
     3661 gcggtctcag atgctgagac acatgaggct gagcaagaca gagagggaca gggaggcaca
     3721 gctggtgaag gatagaaaca gcatgctgag gctgaccagc atcggctccg acgaggatga
     3781 ggagacagag acctatcagg agaaggtgca catgacatgg accaaggaca agtacatggc
     3841 ctcccgcggc cagaaggcca agtctatgga gggcttccag gacctgctga acatgcggcc
     3901 cgatcagagc aatgtgcgga gaatgcacac cgccgtgaag ctgaacgaag tgatcgtgaa
     3961 taagtcccac gaggccaagc tggtgctgct gaatatgcct ggaccaccaa ggaacccaga
     4021 gggcgatgag aattatatgg agtttctgga ggtgctgaca gagggcctgg agcgcgtgct
     4081 gctggtgcgg ggaggaggaa gcgaagtgat caccatctac tccgacccag ctttcttgta
     4141 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     4201 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     4261 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     4321 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     4381 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     4441 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     4501 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     4561 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     4621 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     4681 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     4741 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     4801 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     4861 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     4921 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     4981 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     5041 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     5101 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     5161 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     5221 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     5281 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
