LOCUS       sequence-219921-        8179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ATB0,+.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8179)
  AUTHORS   Samluk L, Czeredys M, Skowronek K, Nalecz KA
  TITLE     Protein kinase C regulates amino acid transporter ATB(0,+).
  JOURNAL   Biochem Biophys Res Commun. 2012 May 25;422(1):64-9. doi: 
            10.1016/j.bbrc.2012.04.106. Epub 2012 Apr 27.
  PUBMED    22561015
REFERENCE   2  (bases 1 to 8179)
  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: 
            "10.1016/j.bbrc.2012.04.106"; journalName: "Biochem Biophys Res 
            Commun"; date: "2012-05-25- 25"; volume: "422"; issue: "1"; pages: 
            "64-9"
COMMENT     Sequence Label: p3 FLAG_CMV14/B0,+
FEATURES             Location/Qualifiers
     source          1..8179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2545..2610
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2621..3243
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2820..2839)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3267..3287)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3285..3582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3636..4430
                     /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(3690..3709)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4300..4319
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5082..5216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5119..5138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5173..5192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5221..5240)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5222..5240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5254..5270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5254..5270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5267..5289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5278..5294
                     /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)."
     promoter        complement(join(5302..5308,5309..5326,5327..5332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5347..5368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5485..5502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5656..6244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5736..5755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6415..7206,7207..7275))
                     /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     7038..7057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7276..7380)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7407..7862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7494..7513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7704..7725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7988..8010
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8003..8019
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaagc ttatggacag attgaagtgc ccgagcttct tcaagtgcag acagaaggag
      661 aaagtgacag cttcatctga gaacttccat gttggtgaaa atgatgaaaa tcaggagcgt
      721 ggtaactggt ccaaaaaatc agattatctt ctttccatgg ttggctatgc ggtgggattg
      781 ggcaacgtgt ggcgattccc ttacctgacc tacaccaatg gtggaggtgc tttcttgata
      841 ccctatgcaa ttatgctagc attggctggc ttgcctctgt tcttccttga atgttcactg
      901 ggacaatttg ctagcttagg tccagtttca gtttggagga ttttgccact gtttcaaggt
      961 gtgggaatca cgatggtcct gatatccatt tttgtgacaa tttattataa tgtcatcatt
     1021 gcctatagtc tttactactt atttgcttct tttcaaagtg tactgccatg ggctaattgt
     1081 tcttcttctt gggcagacga taactgtagc agacaaccaa tagtaactgg ctgtcgtgtg
     1141 agcacagaca caggaggaga catgtttatg aatataagct gggtaaatag caacaacttc
     1201 acctgcttga atggtagtga agtttttcgg ccaggacaac ttcccagtga acagtattgg
     1261 gataaagtga cgcttcggag gtcaagtgga atggatgaaa ctggcgtaat tgtgtggtac
     1321 ttagcacttt gccttcttct agcttggatc atagttggag cagcactatt taaaggaatc
     1381 aaatcctctg gcaaggtggt atattttaca gctcttttcc cgtatgtggt ccttctcatt
     1441 ctactaatac gaggtgcaac tttggagggt gcatcaaaag gcatttcata ttacattgga
     1501 gcacagtcaa attttacaaa actcagagaa gctgaggttt ggaaggatgc tgccactcag
     1561 atattttatt ccctttctgt ggcttggggt ggcctagtgg ctctgtcatc ttacaacaag
     1621 ttcaacaaca actgttactc tgatgccatt atagtttgct cgaccaactg cctcactagt
     1681 gtgtttgctg gattcgccat tttttctatt ctgggacaca tggctcatat atccggaaag
     1741 gaagtctctc aagttgtaaa atcaggattt gacttggcat tcattgctta tccggaagca
     1801 ctagctcaac tcccaggtgg tccattctgg tccatattat ttttcttcat gcttttaaca
     1861 ttgggtctgg attctcaatt tgcttctatt gaaacaatca caacaacaat tcaagatttg
     1921 tttcccaaag caatgaaacg catgagggtt cctataactt tgggatgctg cttgattttg
     1981 tttctccttg gtctcgtctg tgtgactcag gctggaattt actgggttca tttgattgat
     2041 cacttctgtg ctggatgggg gatcttaatt gctgccatct tggaaattgc aggaatcatc
     2101 tggatttatg gaggaaatag gttcattgaa gatatagaaa tgatgattgg agcaaaaaga
     2161 tggctattct ggctttggtg gagagcttgt tggtttgtca tcactcctat actgttgtct
     2221 gcaattctag tctggtcact ggtgaaattt cacagaccta attatgctga tattccgtat
     2281 cctgactggg gagtcgctct cggatggtgt atgattattt tctgcattat ttggataccg
     2341 attatggcga tcataaaaat agttcgggct gaaggaaaca tccttcagcg tatcataaaa
     2401 tgctgtagac cagcttcaaa ctggggtcca tacctggaaa aacatcgtgg agagagatat
     2461 aaagacatgg ctgaacctgc aaaggagact gaccacgaaa tacctactat tagtggcact
     2521 agaaaaccag aaggatcccg ggctgactac aaagaccatg acggtgatta taaagatcat
     2581 gacatcgact acaaggatga cgatgacaag tagtgatccc gggtggcatc cctgtgaccc
     2641 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2701 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2761 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2821 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2881 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     2941 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3001 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3061 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
     3121 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
     3181 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
     3241 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3361 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3421 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3481 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3541 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggaattg atcagcttgg
     3601 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3661 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3721 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3781 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3841 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3901 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3961 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4021 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4081 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4141 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4201 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4261 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4321 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4381 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4441 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4501 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4561 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccgggctcg atcccctcgc
     4621 gagttggttc agctgctgcc tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa
     4681 atccgtcggc atccaggaaa ccagcagcgg ctatccgcgc atccatgccc ccgaactgca
     4741 ggagtgggga ggcacgatgg ccgctttggt cgacccggac gggacgctcc tgcgcctgat
     4801 acagaacgaa ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc
     4861 actgcgtgga tggagcgctg gcgcctgctg cgcgacggcg agctgctcac cacccactcg
     4921 ccaagctgga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccgatca
     4981 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5041 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaatt
     5221 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     5281 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5341 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5401 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5461 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5521 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5581 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5641 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5701 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5761 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5821 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5881 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5941 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6001 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6061 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6121 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6181 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6241 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6301 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6361 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6421 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6481 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6541 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6601 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6661 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6721 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6781 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6841 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6901 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6961 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7021 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7081 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7141 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7201 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7261 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7321 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7381 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     7441 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     7501 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     7561 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     7621 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     7681 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     7741 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     7801 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     7861 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7921 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7981 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctatacattg
     8041 aatcaatatt ggcaattagc catattagtc attggttata tagcataaat caatattggc
     8101 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     8161 ccaatatgac cgccatgtt
//
