LOCUS       sequence-444192-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SLC17A1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  TITLE     SLC17 transporters mediate renal excretion of Lac-Phe in mice and 
            humans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6851)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCDNA3.SLC17A1-flag
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      332..920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     821..840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1074..1091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1208..1229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1244..1249,1250..1267,1268..1274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1282..1298
                     /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     1287..1309
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1306..1322
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1306..1322
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1371..1492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1382..1401)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1436..1455
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1666..2460)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1777..1796)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2387..2406
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2527..2856)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2541..2676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2595..2614)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2841..2861
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2870..3298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2980..3001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3192..3211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3344..3568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3557..3574
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3594..3612
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3594..3611
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3652..3675)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5134..5153)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5135..5153)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5198..5401)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5227..5247)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5402..5781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5953..5972
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6047..6151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6152..6220,6221..6851,1..161)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6370..6389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
       61 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      121 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      661 tgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1081 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1141 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1201 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1261 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1321 accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat
     1381 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1441 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg
     1501 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     1561 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     1621 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     1681 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     1741 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     1801 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     1861 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     1921 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     1981 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     2041 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     2101 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     2161 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     2221 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     2281 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     2341 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     2401 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     2461 gcgaaacgat cctcatcctg tctcttgatc agatccgaaa atggatatac aagctcccgg
     2521 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     2581 agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     2641 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     2701 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     2761 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     2821 ggagcctggg gactttccac accctaactg acacacattc cacagaatta attcgcgtta
     2881 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2941 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3001 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3061 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3121 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3181 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3241 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg
     3301 ggataccccc tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca
     3361 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     3421 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     3481 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     3541 acagatggct ggcaactaga aggcacagtc gaggctgatc agcgagctct agcatttagg
     3601 tgacactata gaatagggcc ctctagatgc atgctcgagc catcattaaa ccttatcgtc
     3661 gtcatccttg taatcactgc cagatccgcc ggcgcgcctt tagagacgtg tgtgttgttt
     3721 ttctttagcc cagtcctgaa tttctgctgt agcaactata aggtagaaaa ttaggccagt
     3781 cacattaatg gctgccatca ggatgaaggt tttaaaccag gcggattccg gatcctgctt
     3841 aaggatcaat ccagtcaaag tggaagcaat tagtcctcct atcattccag ttaaagttga
     3901 acatgcttta ataaatccaa aatatctggg agcaatatcc aagccattta taaacactcc
     3961 acccaagcaa aagctgcctg ttgcaccagc aagtattagg aaaatgacaa tgctgtagaa
     4021 ggtggaactc aggtaaggca ggcagacacc aaagattgca ggaaggagaa atcctgctgc
     4081 tgtgaagagt ttccggacag caattacgct gagaatattc ctggtcagga agaagtctga
     4141 taactgacct gctaggttac cacagatcca ggcaaacaaa tagggaaggg aagacaagaa
     4201 cccattctct tttatattaa catgaagcat ggagttgata aacattggag tgtatagtgt
     4261 catgatgtta tgtgaccaga aaaacgtaaa actaccagtg gaaatagccc agactggaag
     4321 cgacttaagt atagccttga taggcagaga ttgtctactt gaactgacct gctggaccag
     4381 ggaggatgtg atgtattcct tttcactgat gcttatacat gggtggtctt tggggtcatc
     4441 ataaaacaga acgaaccaga gaagacatac ggcacagcca caagcaccaa aaatatagaa
     4501 gaccatgggc cagcccagag attcacagat aactccagtc acaagtagga caataaaggg
     4561 tcccagcaaa aaccctgatg tactcataga agtaagtcgg cctcgttcca ggggaggagc
     4621 ccatttgaca tatatttcaa actgggctgt tgcaactatc ccctgggctg ctccctgaac
     4681 tgctcgacat acaacgaccc aagctactcc aattccagct gctggtggga tgagcaggct
     4741 taacacagag ctgaggcata atgcaaagcc aatcattttc tttgtagaat atattccaga
     4801 gaagtatcca acaggaactt ggatgatgat gacaccatag gaggtggaac tcaagatgat
     4861 tccctggata tctgggctcc aattatacat agggttcttt atattatcca ggagcttctt
     4921 tgtggaggtg ttgggcaaac catgtggatc tgtgctattc accatgacta ccattgtgag
     4981 gttcaggcac gcacgctgtg ctgttattat aacattacaa cagtgcacaa ggaaagacaa
     5041 tccatagcga aaggaacaga aacctggaac ttttttggga ggcaaccggt tatccatttg
     5101 catggtggcg gccgcctaga taagcttggg tctccctata gtgagtcgta ttaatttcga
     5161 taagccagta agcagtgggt tctctagtta gccagagagc tctgcttata tagacctccc
     5221 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     5281 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     5341 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     5401 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     5461 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     5521 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     5581 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     5641 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     5701 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     5761 aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa
     5821 aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag
     5881 cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact
     5941 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag
     6001 atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6061 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6121 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6181 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6241 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6301 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6361 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6421 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6481 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6541 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6601 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6661 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6721 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6781 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6841 tgataaatct g
//
