LOCUS       sequence-435706-        4625 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 4625)
  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 4625)
  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-SLC17A1_STOP
FEATURES             Location/Qualifiers
     source          1..4625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        298..402
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(406..471,472..1263)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="SIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIE
                     LDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSP
                     VTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWE
                     PELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSAL
                     PAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASL
                     IKHW"
     primer_bind     complement(621..640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1434..2022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1923..1942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2310..2331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2346..2351,2352..2369,2370..2376)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2384..2400
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2389..2411
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2408..2424
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2408..2424
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2473..2594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2484..2503)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2538..2557
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2717..3430,3431..3433,3434..3436))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2720..3430,3431..3433,3434..3436))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2762..2783)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3111..3130
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3370..3391
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3433..3442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3728..3931)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3757..3777)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     repeat_region   4241..4370
                     /label=AAV2 ITR
     rep_origin      join(4442..4625,1..272)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4529..4548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
       61 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      121 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      181 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      241 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
      301 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      361 taaccctgat aaatgcttca ataatattga aaaaggaaga gtacgagtat tcaacatttc
      421 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      481 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      541 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      601 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      661 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      721 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      781 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      841 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      901 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      961 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1021 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1081 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1141 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1201 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1261 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1321 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1381 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1441 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1501 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1561 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1621 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1681 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1741 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1801 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1861 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1921 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1981 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2041 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2101 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2161 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2221 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2281 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2341 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2401 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt
     2461 atacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2521 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2581 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     2641 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     2701 tagagtcgcg gccgctttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     2761 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     2821 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     2881 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     2941 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     3001 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3061 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3121 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3181 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3241 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     3301 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     3361 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     3421 cgcccttgct caccatggtg gcgatggatc cggaagagta ggcgcgcttc tggaaagcct
     3481 ggcgggtggt ggcgttgacg atggaggtca tctgggtgcg cttgagggtc tggagggggg
     3541 agccagtctg gatggtgcgc ttgctgacct gagcaacgcg gacagcaggg cgggcaacct
     3601 tggcggaagc agccatccgg gaggccaggc gagaggcgag gacacgagtg gaggccatgg
     3661 tgaattcgaa gcttgagctc gagatctgag tccggtagcg ctagcggatc tgacggttca
     3721 ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     3781 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     3841 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     3901 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     3961 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4021 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     4081 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     4141 gcgttactat gggaacatac gtcattattg acgtcaatgg tactcttcct agagcatggc
     4201 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     4261 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaagggcgcc
     4321 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat
     4381 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4441 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4501 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4561 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4621 agtgc
//
