LOCUS       sequence-365731-        3597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3597)
  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 3597)
  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-SLC10A2_STOP
FEATURES             Location/Qualifiers
     source          1..3597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1154..1253)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1270..1289)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1271..1289)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1294..1310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1294..1310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1412..1431)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1423..2232
                     /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(1490..1509)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2379..2967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2868..2887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3121..3138
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3297..3324)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3416..3502)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3551..3573
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3565..3582
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3566..3582
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgaa cgatcccaat
      121 tcttgcgtgg acaacgcaac cgtgtgcagc ggagcatcct gcgtggtgcc tgagagcaat
      181 ttcaacaata tcctgtccgt ggtgctgtct accgtgctga caatcctgct ggccctggtc
      241 atgttctcca tgggctgtaa cgtggagatc aagaagtttc tgggccacat caagaggcct
      301 tggggcatct gcgtgggctt cctgtgccag tttggcatca tgccactgac cggcttcatc
      361 ctgtctgtgg cctttgatat cctgccactg caggccgtgg tggtgctgat catcggatgc
      421 tgtccaggag gaacagcaag caatatcctg gcctactggg tggacggcga tatggacctg
      481 tctgtgagca tgaccacatg ctccaccctg ctggccctgg gcatgatgcc actgtgcctg
      541 ctgatctaca caaagatgtg ggtggattcc ggctctatcg tgatccccta tgacaacatc
      601 ggcacctctc tggtgagcct ggtggtgccc gtgagcatcg gcatgtttgt gaatcacaag
      661 tggcctcaga aggccaagat catcctgaag atcggctcca tcgccggcgc catcctgatc
      721 gtgctgatcg ccgtggtggg cggcatcctg tatcagtctg cctggatcat cgccccaaag
      781 ctgtggatca tcggcacaat cttccccgtg gccggctact ccctgggctt tctgctggca
      841 aggatcgcag gactgccttg gtatcggtgc agaaccgtgg ccttcgagac aggcatgcag
      901 aacacccagc tgtgcagcac aatcgtgcag ctgtccttta cccctgagga gctgaatgtg
      961 gtgttcacat ttccactgat ctactctatc ttccagctgg cctttgccgc catcttcctg
     1021 ggcttttacg tggcctataa gaagtgccac ggcaagaaca aggccgagat ccccgagagc
     1081 aaggagaatg gcaccgagcc tgagagctcc ttctataagg ccaacggcgg ctttcagccc
     1141 gacgagaagt aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1201 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1261 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1321 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1381 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1441 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1501 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1561 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1621 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1681 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1741 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1801 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1861 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1921 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1981 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2041 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2101 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2161 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2221 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2281 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2521 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3001 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3061 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3241 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3301 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3361 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3421 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     3481 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     3541 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
