LOCUS       sequence-8755-ex        7795 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 7795)
  AUTHORS   Takanaga H, Chaudhuri B, Frommer WB
  TITLE     GLUT1 and GLUT9 as major contributors to glucose influx in HepG2 
            cells identified by a high sensitivity intramolecular FRET glucose 
            sensor.
  JOURNAL   Biochim Biophys Acta. 2008 Apr . 1778(4):1091-9.
  PUBMED    18177733
REFERENCE   2  (bases 1 to 7795)
  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: "Biochim 
            Biophys Acta. 2008 Apr . 1778(4):1091-9."
COMMENT     Sequence Label: pcDNA3.1 FLII12Pglu-700uDelta6
FEATURES             Location/Qualifiers
     source          1..7795
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1032..1034,1035..1037,1038..1745)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(2643..2645,2646..2648,2649..3359)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3389..3406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3395..3619
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3665..4093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3752..3771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3962..3983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4102..4122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4107..4436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4287..4422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4349..4368
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4503..5297
                     /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(4557..4576)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5167..5186
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5471..5592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5508..5527)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5562..5581
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5641..5657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5641..5657)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5654..5676)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5665..5681
                     /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(5689..5695,5696..5713,5714..5719))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5734..5755
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5872..5889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6043..6628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6123..6142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6799..7590,7591..7659))
                     /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     7422..7441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7660..7764)
                     /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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtggat ccgaccatgg ctgatactcg cattggtgta acaatctata
     1021 agtcggctgg tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1081 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1141 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1201 cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc cgctaccccg
     1261 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1321 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1381 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1441 tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc accgccgaca
     1501 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg
     1561 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1621 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1681 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1741 tatacggtag ccgagataac tttatgtctg tagtgcgcaa ggctattgag caagatgcga
     1801 aagccgcgcc agatgttcag ctgctgatga atgattctca gaatgaccag tccaagcaga
     1861 acgatcagat cgacgtattg ctggcgaaag gggtgaaggc actggcaatc aacctggttg
     1921 acccggcagc tgcgggtacg gtgattgaga aagcgcgtgg gcaaaacgtg ccggtggttt
     1981 tcttcaacaa agaaccgtct cgtaaggcgc tggatagcta cgacaaagcc tactacgttg
     2041 gcactgactc caaagagtcc ggcattattc aaggcgattt gattgctaaa cactgggcgg
     2101 cgaatcaggg ttgggatctg aacaaagacg gtcagattca gttcgtactg ctgaaaggtg
     2161 aaccgggcca tccggatgca gaagcacgta ccacttacgt gattaaagaa ttgaacgata
     2221 aaggcatcaa aactgaacag ttacagttag ataccgcaat gtgggacacc gctcaggcga
     2281 aagataagat ggacgcctgg ctgtctggcc cgaacgccaa caaaatcgaa gtggttatcg
     2341 ccaacaacga tgcgatggca atgggcgcgg ttgaagcgct gaaagcacac aacaagtcca
     2401 gcattccggt gtttggcgtc gatgcgctgc cagaagcgct ggcgctggtg aaatccggtg
     2461 cactggcggg caccgtactg aacgatgcta acaaccaggc gaaagcgacc tttgatctgg
     2521 cgaaaaacct ggccgatggt aaaggtgcgg ctgatggcac caactggaaa atcgacaaca
     2581 aagtggtccg cgtaccttat gttggcgtag ataaagacaa cctggctgaa ttcagcaaga
     2641 aaatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2761 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2821 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3061 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3121 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3181 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3241 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3301 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3361 aaaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3421 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3481 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3541 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3601 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3661 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3721 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3781 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3841 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3901 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3961 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4021 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4081 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4201 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4261 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4321 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4381 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4441 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4501 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4561 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4621 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4681 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4741 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4801 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4861 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4921 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4981 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5041 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5101 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5161 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5221 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5281 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5341 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5401 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5461 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5521 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5581 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5641 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5701 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5761 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5821 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5881 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5941 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6481 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
