LOCUS       sequence-8774-ex        7870 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7870)
  AUTHORS   Takanaga H, Frommer WB
  TITLE     Facilitative plasma membrane transporters function during ER 
            transit.
  JOURNAL   FASEB J. 2010 Aug . 24(8):2849-58.
  PUBMED    20354141
REFERENCE   2  (bases 1 to 7870)
  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"; journalName: "FASEB J. 
            2010 Aug . 24(8):2849-58."
COMMENT     Sequence Label: pcDNA3.1 FLIPglu-170nDelta13V
FEATURES             Location/Qualifiers
     source          1..7870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(942..944,945..947,948..1460)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             2994..3245
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     protein_bind    complement(3260..3284)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3347..3365)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3464..3481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3470..3694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3740..4168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4177..4197)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4182..4511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4362..4497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4424..4443
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4578..5372
                     /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(4632..4651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5242..5261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5546..5667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5583..5602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5637..5656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5716..5732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5716..5732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5729..5751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5740..5756
                     /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(5764..5770,5771..5788,5789..5794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5809..5830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5947..5964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6118..6703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6198..6217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6874..7665,7666..7734))
                     /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     7497..7516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7735..7839)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgggccg catggtgagc aagggcgagg
      961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1081 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1141 ggggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1381 tcagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1441 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1561 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1621 ccgccgggat cgctgatact cgcattggtg taacaatcta taagtacgac gataacttta
     1681 tgtctgtagt gcgcaaggct attgagcaag atgcgaaagc cgcgccagat gttcagctgc
     1741 tgatgaatga ttctcagaat gaccagtcca agcagaacga tcagatcgac gtattgctgg
     1801 cgaaaggggt gaaggcactg gcaatcaacc tggttgaccc ggcagctgcg ggtacggtga
     1861 ttgagaaagc gcgtgggcaa aacgtgccgg tggttttctt caacaaagaa ccgtctcgta
     1921 aggcgctgga tagctacgac aaagcctact acgttggcac tgactccaaa gagtccggca
     1981 ttattcaagg cgatttgatt gctaaacact gggcggcgaa tcagggttgg gatctgaaca
     2041 aagacggtca gattcagttc gtactgctga aaggtgaacc gggccatccg gatgcagaag
     2101 cacgtaccac ttacgtgatt aaagaattga acgataaagg catcaaaact gaacagttac
     2161 agttagatac cgcaatgtgg gacaccgctc aggcgaaaga taagatggac gcctggctgt
     2221 ctggcccgaa cgccaacaaa atcgaagtgg ttatcgccaa caacgatgcg atggcaatgg
     2281 gcgcggttga agcgctgaaa gcacacaaca agtccagcat tccggtgttt ggcgtcgatg
     2341 cgctgccaga agcgctggcg ctggtgaaat ccggtgcact ggcgggcacc gtactgaacg
     2401 atgctaacaa ccaggcgaaa gcgacctttg atctggcgaa aaacctggcc gatggtaaag
     2461 gtgcggctga tggcaccaac tggaaaatcg acaacaaagt ggtccgcgta ccttatgttg
     2521 gcgtagataa agacaacctg gctctcgagc tgttcaccgg ggtggtgccc atcctggtcg
     2581 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2641 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     2701 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
     2761 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2821 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2881 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2941 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     3001 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     3061 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3121 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3181 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3241 acaagtaaaa gctatctaga cccagctttc ttgtacaaag tggtgtagtt aattcatgat
     3301 ggccgctgca ggtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     3361 tattacgcgc ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg
     3421 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3481 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3541 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3601 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3661 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3721 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4261 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4381 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4441 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4501 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4561 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4621 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4681 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4741 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4801 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4861 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4921 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4981 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5041 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5101 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5161 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5221 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5281 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5341 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5401 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5461 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5521 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5641 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5701 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5761 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5821 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6541 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6721 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6841 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6901 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6961 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7021 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7081 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7141 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7201 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7261 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7321 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7381 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7441 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7501 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7561 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7621 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7681 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7741 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7801 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7861 acctgacgtc
//
