LOCUS       sequence-342380-        4128 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 4128)
  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 4128)
  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-SLC18A1_STOP
FEATURES             Location/Qualifiers
     source          1..4128
                     /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"
     RBS             267..275
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(1685..1784)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1801..1820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1802..1820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1825..1841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1825..1841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1943..1962)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1954..2763
                     /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(2021..2040)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2910..3498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3399..3418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3652..3669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3828..3855)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3947..4033)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4082..4104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4096..4113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4097..4113
                     /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 ccaccatgct gaggaccatc
      121 ctggacgcac cacagagact gctgaaggag ggaagggcaa gcaggcagct ggtgctggtg
      181 gtggtgttcg tggccctgct gctggacaat atgctgttta ccgtggtggt gccaatcgtg
      241 cccacattcc tgtacgatat ggagtttaag gaggtgaaca gctccctgca cctgggacac
      301 gcaggatcta gcccacacgc actggcatct cctgccttca gcaccatctt ttccttcttt
      361 aacaataaca cagtggcagt ggaggagagc gtgccatccg gaatcgcctg gatgaatgat
      421 accgcatcca caatcccacc tccagcaacc gaggcaatct ctgcccacaa gaataactgc
      481 ctgcagggaa ccggcttcct ggaggaggag atcacacggg tgggcgtgct gtttgccagc
      541 aaggccgtga tgcagctgct ggtgaatccc ttcgtgggcc ctctgacaaa cagaatcggc
      601 taccacatcc ccatgttcgc cggctttgtg atcatgtttc tgtccaccgt gatgttcgcc
      661 ttttctggca cctatacact gctgttcgtg gcacggacac tgcagggaat cggatcctct
      721 tttagctccg tggcaggact gggaatgctg gcatccgtgt acaccgacga tcacgagcgg
      781 ggcagagcca tgggaacagc cctgggagga ctggccctgg gcctgctggt gggcgcccct
      841 ttcggcagcg tgatgtatga gtttgtgggc aagtccgccc cattcctgat cctggccttt
      901 ctggccctgc tggacggcgc cctgcagctg tgcatcctgc agccatctaa ggtgagccca
      961 gagtccgcca agggaacccc actgttcatg ctgctgaagg atccctacat cctggtggcc
     1021 gccggctcta tctgctttgc caatatggga gtggcaatcc tggagcctac cctgccaatc
     1081 tggatgatgc agacaatgtg cagccctaag tggcagctgg gactggcctt cctgccagca
     1141 tccgtgtctt atctgatcgg caccaatctg tttggcgtgc tggccaacaa gatgggcagg
     1201 tggctgtgct ctctgatcgg catgctggtg gtgggcacaa gcctgctgtg cgtgccactg
     1261 gcccacaata tcttcggact gatcggacct aacgcaggac tgggactggc aatcggaatg
     1321 gtggactcta gcatgatgcc tatcatgggc cacctggtgg atctgcgcca caccagcgtg
     1381 tacggcagcg tgtatgccat cgccgacgtg gccttctgca tgggctttgc aatcggacca
     1441 tccaccggag gagcaatcgt gaaggcaatc ggcttcccct ggctgatggt catcacaggc
     1501 gtgatcaaca tcgtgtacgc ccccctgtgc tactatctga ggagcccccc tgccaaggag
     1561 gagaagctgg ccatcctgtc ccaggactgt cctatggaga cacgcatgta tgccacccag
     1621 aagcctacaa aggagtttcc actgggcgag gactctgatg aggagcccga tcacgaggag
     1681 taagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     1741 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     1801 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1861 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1921 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1981 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2041 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2101 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2161 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2221 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2281 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2341 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2401 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2461 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2521 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2581 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2641 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2701 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2761 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     2821 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2881 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2941 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3001 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3061 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3121 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3181 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3241 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3301 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3361 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3421 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3481 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3541 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3601 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3661 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3721 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3781 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     3841 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     3901 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     3961 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     4021 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     4081 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
