LOCUS       sequence-147541-        7638 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 7638)
  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 7638)
  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: GLUT1-eGFP/pcDNA-DEST47
FEATURES             Location/Qualifiers
     source          1..7638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2033..2052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2224..2603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2604..2807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2758..2778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2852..2871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2852..2870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2903..2927
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4440..4464)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4475..4477,4478..4480,4481..5194)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4508..4536)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5131..5151
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5216..5233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5222..5446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5492..5920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5579..5598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5789..5810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5929..5949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5934..6263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6114..6249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6176..6195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6330..7124
                     /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(6384..6403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6994..7013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7300..7421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7337..7356)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7391..7410
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7470..7486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7470..7486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7483..7505)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7494..7510
                     /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(7518..7524,7525..7542,7543..7548))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7563..7584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     2881 ctggctagtt aagcttgatc aaacaagttt gtacaaaaaa gcaggctccg cggccgcccc
     2941 cttcaccatg gagcccagca gcaagaagct gacgggtcgc ctcatgctgg ccgtgggagg
     3001 agcagtgctt ggctccctgc agtttggcta caacactgga gtcatcaatg ccccccagaa
     3061 ggtgatcgag gagttctaca accagacatg ggtccaccgc tatggggaga gcatcctgcc
     3121 caccacgctc accacgctct ggtccctctc agtggccatc ttttctgttg ggggcatgat
     3181 tggctccttc tctgtgggcc ttttcgttaa ccgctttggc cggcggaatt caatgctgat
     3241 gatgaacctg ctggccttcg tgtccgccgt gctcatgggc ttctcgaaac tgggcaagtc
     3301 ctttgagatg ctgatcctgg gccgcttcat catcggtgtg tactgcggcc tgaccacagg
     3361 cttcgtgccc atgtatgtgg gtgaagtgtc acccacagcc cttcgtgggg ccctgggcac
     3421 cctgcaccag ctgggcatcg tcgtcggcat cctcatcgcc caggtgttcg gcctggactc
     3481 catcatgggc aacaaggacc tgtggcccct gctgctgagc atcatcttca tcccggccct
     3541 gctgcagtgc atcgtgctgc ccttctgccc cgagagtccc cgcttcctgc tcatcaaccg
     3601 caacgaggag aaccgggcca agagtgtgct aaagaagctg cgcgggacag ctgacgtgac
     3661 ccatgacctg caggagatga aggaagagag tcggcagatg atgcgggaga agaaggtcac
     3721 catcctggag ctgttccgct cccccgccta ccgccagccc atcctcatcg ctgtggtgct
     3781 gcagctgtcc cagcagctgt ctggcatcaa cgctgtcttc tattactcca cgagcatctt
     3841 cgagaaggcg ggggtgcagc agcctgtgta tgccaccatt ggctccggta tcgtcaacac
     3901 ggccttcact gtcgtgtcgc tgtttgtggt ggagcgagca ggccggcgga ccctgcacct
     3961 cataggcctc gctggcatgg cgggttgtgc catactcatg accatcgcgc tagcactgct
     4021 ggagcagcta ccctggatgt cctatctgag catcgtggcc atctttggct ttgtggcctt
     4081 ctttgaagtg ggtcctggcc ccatcccatg gttcatcgtg gctgaactct tcagccaggg
     4141 tccacgtcca gctgccattg ccgttgcagg cttctccaac tggacctcaa atttcattgt
     4201 gggcatgtgc ttccagtatg tggagcaact gtgtggtccc tacgtcttca tcatcttcac
     4261 tgtgctcctg gttctgttct tcatcttcac ctacttcaaa gttcctgaga ctaaaggccg
     4321 gaccttcgat gagatcgctt ccggcttccg gcagggggga gccagccaaa gtgacaagac
     4381 acccgaggag ctgttccatc ccctgggggc tgattcccaa gtgaagggtg ggcgcgccga
     4441 cccagctttc ttgtacaaag tggttcgatc tagaatggct agcaaaggag aagaactttt
     4501 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     4561 tgtcagtgga gagggtgaag gtgatgctac atacggaaag cttaccctta aatttatttg
     4621 cactactgga aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt
     4681 tcaatgcttt tcccgttatc cggatcatat gaaacggcat gactttttca agagtgccat
     4741 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac
     4801 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat
     4861 tgattttaaa gaagatggaa acattctcgg acacaaactc gagtacaact ataactcaca
     4921 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     4981 ccacaacatt gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat
     5041 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc
     5101 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     5161 gattacacat ggcatggatg agctctacaa ataatgaatt aaacccgctg atcagcctcg
     5221 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5281 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5341 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5401 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     5461 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     5521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5641 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6481 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6841 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     7021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     7141 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     7201 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7261 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7321 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7381 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7441 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7501 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7561 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7621 aacctgtcgt gccagctg
//
