LOCUS       sequence-260963-        4344 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 4344)
  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 4344)
  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_SLC23A1
FEATURES             Location/Qualifiers
     source          1..4344
                     /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(2469..2568)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2585..2604)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2586..2604)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2609..2625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2609..2625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2727..2746)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2738..3547
                     /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(2805..2824)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3694..4282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4183..4202
                     /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 accatgaggg cacaggagga cctggaggga aggacccagc acgagaccac
      721 acgcgatcca agcacaccac tgcccaccga gcccaagttc gacatgctgt acaagatcga
      781 ggatgtgccc ccttggtatc tgtgcatcct gctgggcttc cagcactacc tgacatgttt
      841 ttctggcacc atcgcagtgc cctttctgct ggcagaggcc ctgtgcgtgg gacacgacca
      901 gcacatggtg agccagctga tcggcacaat cttcacctgc gtgggcatca ccacactgat
      961 ccagaccaca gtgggcatcc ggctgcccct gtttcaggcc tccgccttcg cctttctggt
     1021 gcctgccaag gccatcctgg ccctggagag atggaagtgc ccacccgagg aggagatcta
     1081 cggcaattgg tctctgcctc tgaacaccag ccacatctgg cacccaagga tcagggaggt
     1141 gcagggagcc atcatggtga gctccgtggt ggaggtggtc atcggactgc tgggactgcc
     1201 tggcgccctg ctgaattata tcggccctct gaccgtgaca ccaaccgtga gcctgatcgg
     1261 actgtccgtg ttccaggcag caggcgatag ggcaggatcc cactggggaa tctccgcctg
     1321 ctctatcctg ctgatcatcc tgttctctca gtacctgcgc aacctgacat ttctgctgcc
     1381 cgtgtacaga tggggcaagg gactgaccct gctgagaatc cagatcttca agatgtttcc
     1441 aatcatgctg gccatcatga ccgtgtggct gctgtgctac gtgctgacac tgaccgacgt
     1501 gctgccaaca gatcccaagg cctatggatt tcaggcaagg accgacgcaa gaggcgatat
     1561 catggccatc gccccatgga tcaggatccc ttacccatgc cagtggggac tgccaacagt
     1621 gaccgcagca gccgtgctgg gaatgttcag cgccaccctg gcaggcatca tcgagtccat
     1681 cggcgactac tatgcctgtg caaggctggc aggagcacct ccaccacctg tgcacgccat
     1741 caatcgcggc atctttacag agggcatctg ctgtatcatc gcaggactgc tgggaacagg
     1801 aaatggaagc acctctagct cccccaacat cggcgtgctg ggcatcacca aagtgggatc
     1861 caggagagtg gtgcagtacg gagcagcaat catgctggtg ctgggaacaa tcggcaagtt
     1921 caccgccctg ttttctagcc tgcccgaccc tatcctgggc ggcatgttct gcacactgtt
     1981 tggcatgatc accgccgtgg gcctgtccaa cctgcagttc gtggatatga actcctctcg
     2041 caatctgttc gtgctgggct tttctatgtt ctttggcctg acactgccca actatctgga
     2101 gagcaatcct ggcgccatca acaccggcat cctggaggtg gaccagatcc tgatcgtgct
     2161 gctgaccaca gagatgttcg tgggcggctg tctggccttt atcctggata atacagtgcc
     2221 tggatctcca gaggagaggg gactgatcca gtggaaggca ggagcacacg caaactccga
     2281 catgagctcc tctctgaagt cttacgattt ccccatcggc atgggcatcg tgaagagaat
     2341 cacctttctg aagtatatcc ccatctgccc cgtgttcaag ggctttagct cctctagcaa
     2401 ggaccagatc gccatcccag aggatacacc cgagaataca gagaccgcca gcgtgtgcac
     2461 caaggtggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2521 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2581 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2641 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2701 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2761 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2821 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2881 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2941 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3001 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3061 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3121 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3181 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3241 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3301 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3361 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3421 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3481 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3541 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3601 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3841 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4321 ttgctggcct tttgctcaca tgtt
//
