LOCUS       sequence-8777-ex        7904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7904)
  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 7904)
  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: "FASEB J. 
            2010 Aug . 24(8):2849-58."
COMMENT     Sequence Label: pEF/myc/ER FLIPglu-30uDelta13V
FEATURES             Location/Qualifiers
     source          1..7904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..1185
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          254..1176
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1133..1153
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1193..1202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(1199..1243,1326..1337)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(1199..1244,1327..1337)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(1374..1376,1377..1889)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIE"
     CDS             3423..3674
                     /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"
     CDS             3684..3713
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3767..3784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3773..3997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4043..4471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4130..4149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4340..4361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4480..4500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4485..4815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4666..4801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4728..4747
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4836..5630
                     /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(4890..4909)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5500..5519
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5804..5925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5841..5860)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5895..5914
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5956..5973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6127..6715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6207..6226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6886..7677,7678..7746))
                     /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     7509..7528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7747..7851)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt caagcttcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
       61 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      121 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      181 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      241 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      301 tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc ttgatcccga
      361 gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag
      421 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
      481 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
      541 ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat ttcggttttt
      601 gggcccgcgg ccggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
      661 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
      721 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
      781 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa
      841 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
      901 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
      961 ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     1021 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     1081 atgtaattct ccttggaatt tggccttttt gagtttggat cttggttcat tctcaagcct
     1141 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg tggccaccat
     1201 gggatggagc tgtatcatcc tcttcttggt agcaacagct acaggtaagg ggttaacagt
     1261 agcaggcttg aggtctggac atatatatgg gtgacaatga catccacttt gcctttctct
     1321 ccacaggcgc gcactcccag gtccaactgc aggtcgacct cgagagcggc cgcgtgagca
     1381 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1441 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1501 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1561 ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1621 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1681 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1741 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1801 acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1861 ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1921 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1981 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2041 tcgtgaccgc cgccgggatc gctgatactc gcattggtgt aacaatctat aagtacgacg
     2101 ataactttat gtctgtagtg cgcaaggcta ttgagcaaga tgcgaaagcc gcgccagatg
     2161 ttcagctgct gatgaatgat tctcagaatg accagtccaa gcagaacgat cagatcgacg
     2221 tattgctggc gaaaggggtg aaggcactgg caatcaacct ggttgacccg gcagctgcgg
     2281 gtacggtgat tgagaaagcg cgtgggcaaa acgtgccggt ggttttcttc aacaaagaac
     2341 cgtctcgtaa ggcgctggat agctacgaca aagcctacta cgttggcact gactccaaag
     2401 agtccggcat tattcaaggc gatttgattg ctaaacactg ggcggcgaat cagggttggg
     2461 atctgaacaa agacggtcag attcagttcg tactgctgaa aggtgaaccg ggccatccgg
     2521 cggcagaagc acgtaccact tacgtgatta aagaattgaa cgataaaggc atcaaaactg
     2581 aacagttaca gttagatacc gcaatgtggg acaccgctca ggcgaaagat aagatggacg
     2641 cctggctgtc tggcccgaac gccaacaaaa tcgaagtggt tatcgccaac aacgatgcga
     2701 tggcaatggg cgcggttgaa gcgctgaaag cacacaacaa gtccagcatt ccggtgtttg
     2761 gcgtcgatgc gctgccagaa gcgctggcgc tggtgaaatc cggtgcactg gcgggcaccg
     2821 tactgaacga tgctaacaac caggcgaaag cgacctttga tctggcgaaa aacctggccg
     2881 atggtaaagg tgcggctgat ggcaccaact ggaaaatcga caacaaagtg gtccgcgtac
     2941 cttatgttgg cgtagataaa gacaacctgg ctctcgagct gttcaccggg gtggtgccca
     3001 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3061 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     3121 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct
     3181 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3241 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3301 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3361 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     3421 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     3481 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3541 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     3601 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3661 acgagctgta caaggcggcc gcagaacaaa aactcatctc agaagaggat ctgaatgggg
     3721 ccgcaagcga gaaggacgag ctgtagtcta gaagctcgct gatcagcctc gactgtgcct
     3781 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3841 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3901 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3961 aatagcaggc atgctgggga tggcccgggc tctatggctt ctgaggcgga aagaaccagc
     4021 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4081 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4141 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4201 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4261 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4321 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4381 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4441 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4501 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     4561 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4681 ccgcccagtt ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag
     4741 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4801 ctaggctttt gcaaaaagct cccgggaggt ccacaatgat tgaacaagat ggattgcacg
     4861 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4921 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4981 tcaagaccga cctgtccggt gccctgaatg aactccagga cgaggcagcg cggctatcgt
     5041 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5101 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5161 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5221 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5281 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5341 aactgttcgc caggctcaag gcgcgtatgc ccgacggcga ggatctcgtc gtgactcatg
     5401 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5461 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5521 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5581 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5641 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5701 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5761 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5941 ggatctttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6001 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6061 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6301 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6541 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7861 ccgaaaagtg ccacctgacg tcagatcgac ggatcgggag atcg
//
