LOCUS       sequence-263649-        5310 bp    DNA     circular SYN 15-APR-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 5310)
  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 5310)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_SLC14A2
FEATURES             Location/Qualifiers
     source          1..5310
                     /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(3435..3534)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3551..3570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3552..3570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3575..3591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3575..3591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3693..3712)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3704..4513
                     /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(3771..3790)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4660..5248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5149..5168
                     /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 accatgagcg acccacacag ctccccactg ctgccagagc ctctgtctag
      721 cagatacaag ctgtatgagg ccgagtttac ctccccttct tggccaagca cctccccaga
      781 tacacacccc gccctgcctc tgctggagat gcctgaggag aaggacctga ggtcctctaa
      841 cgaggatagc cacatcgtga agatcgagaa gctgaatgag cggagcaagc ggaaggacga
      901 tggagtggca cacagagact ccgccggaca gaggtgcatc tgtctgtcta aggccgtggg
      961 ctacctgacc ggcgatatga aggagtatag aatctggctg aaggacaagc acctggccct
     1021 gcagttcatc gattgggtgc tgaggggcac agcccaggtc atgtttatca acaatcctct
     1081 gtctggcctg atcatcttca tcggcctgct gatccagaac ccatggtgga ccatcacagg
     1141 aggactggga accgtggtgt ccaccctgac agccctggcc ctgggacagg acagatctgc
     1201 catcgccagc ggcctgcacg gctacaatgg catgctggtg ggcctgctga tggccgtgtt
     1261 ttccgagaag ctggattact attggtggct gctgttccca gtgaccttta cagccatgtc
     1321 ttgccccgtg ctgagctccg ccctgaactc catcttctct aagtgggacc tgcccgtgtt
     1381 caccctgccc ttcaacatcg cagtgaccct gtacctggca gcaacaggcc actataatct
     1441 gttctttcct accacactgg tggagcccgt gtctagcgtg cctaacatca cctggacaga
     1501 gatggagatg cctctgctgc tgcaggcaat cccagtggga gtgggacagg tgtacggatg
     1561 cgataatcca tggaccggag gcgtgttcct ggtggccctg tttatctcct ctcctctgat
     1621 ctgtctgcac gccgccatcg gcagcatcgt gggactgctg gccgccctgt ccgtggccac
     1681 cccatttgag accatctaca caggcctgtg gagctataac tgcgtgctgt cctgtatcgc
     1741 catcggcggc atgttttacg ccctgacctg gcagacacac ctgctggccc tgatctgcgc
     1801 cctgttctgt gcctatatgg aggccgccat ctccaatatc atgtctgtgg tgggagtgcc
     1861 acctggaacc tgggcatttt gcctggccac aatcatcttc ctgctgctga ccacaaacaa
     1921 tcccgccatc ttccgcctgc ctctgagcaa ggtgacctac cccgaggcca accggatcta
     1981 ctatctgaca gtgaagtctg gcgaggagga gaaggcacca agcggaggag gaggagagca
     2041 cccacccacc gccggcccca aggtggagga gggaagcgag gccgtgctga gcaagcaccg
     2101 ctccgtgttt cacatcgagt ggagctccat ccggagaagg tccaaggtgt tcggcaaggg
     2161 cgagcaccag gagcggcaga acaaggaccc atttccctac agatatagga agccaaccgt
     2221 ggagctgctg gacctggata caatggagga gtctagcgag atcaaggtgg agaccaatat
     2281 cagcaagaca tcctggatca gatcctctat ggccgccagc ggcaagaggg tgtctaaggc
     2341 cctgagctac atcaccggag agatgaagga gtgcggagag ggcctgaagg acaagtcccc
     2401 cgtgttccag ttctttgatt gggtgctgcg cggcacatct caggtcatgt ttgtgaacaa
     2461 tccactgagc ggcatcctga tcatcctggg cctgttcatc cagaatcctt ggtgggcaat
     2521 ctccggatgt ctgggcacca tcatgtctac cctgacagcc ctgatcctgt cccaggacaa
     2581 gtctgccatc gccgccggct ttcacggcta taatggcgtg ctggtgggac tgctgatggc
     2641 cgtgttcagc gacaagggcg attattactg gtggctgctg ctgcctgtga tcatcatgtc
     2701 tatgagctgc ccaatcctga gctccgccct gggcaccatc ttctccaagt gggatctgcc
     2761 agtgttcaca ctgcccttta acatcaccgt gacactgtac ctggcagcaa ccggacatta
     2821 caatctgttc tttccaacca cactgctgca gcctgcaagc gccatgccaa acatcacatg
     2881 gtccgaggtg caggtgccac tgctgctgag ggcaatccct gtgggaatcg gccaggtgta
     2941 cggctgcgac aatccttgga ccggcggcat cttcctgatc gccctgttta tctctagccc
     3001 actgatttgc ctgcatgctg ccatcggcag cacaatggga atgctggccg ccctgaccat
     3061 cgcaacacct ttcgattcca tctattttgg cctgtgcggc ttcaactcta ccctggcttg
     3121 cattgctatc ggcggcatgt tctacgtgat cacttggcag acccacctgc tggccatcgc
     3181 ctgtgcactg tttgcagcat atctgggagc cgccctggcc aatatgctga gcgtgttcgg
     3241 cctgcctcca tgcacctggc ccttttgtct gagcgccctg acattcctgc tgctgactac
     3301 caacaatcca gccatctaca agctgcccct gagcaaggtg acctatcctg aggccaaccg
     3361 catctactat ctgtctcagg agcggaatcg ccgggccagc atcatcacaa agtaccaggc
     3421 ctatgacgtg agcgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3481 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3541 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     3601 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     3661 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3721 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3781 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3841 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3901 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3961 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     4021 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4081 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4141 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4201 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4261 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4321 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4381 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4441 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4501 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     4561 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4621 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4681 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4741 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4801 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4861 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4921 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4981 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5041 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5101 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5161 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5221 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5281 ggccttttgc tggccttttg ctcacatgtt
//
