LOCUS       sequence-156490-        7825 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7825)
  AUTHORS   Fehr M, Lalonde S, Lager I, Wolff MW, Frommer WB
  TITLE     In vivo imaging of the dynamics of glucose uptake in the cytosol of 
            COS-7 cells by fluorescent nanosensors.
  JOURNAL   J Biol Chem. 2003 May 23. 278(21):19127-33.
  PUBMED    12649277
REFERENCE   2  (bases 1 to 7825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2003 May 23. 278(21):19127-33."
COMMENT     Sequence Label: pcDNA3.1 FLIPglu-600u
FEATURES             Location/Qualifiers
     source          1..7825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2927..2929,2930..2932,2933..3643)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4607..4609,4610..4612,4613..5326)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5353..5370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5359..5583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5629..6057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5716..5735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5926..5947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6066..6086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6071..6400
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6251..6386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6313..6332
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6467..7261
                     /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(6521..6540)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7131..7150
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7435..7556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7472..7491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7526..7545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7605..7621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7605..7621)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7618..7640)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7629..7645
                     /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(7653..7659,7660..7677,7678..7683))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7698..7719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct ggctagcgtt taaacgggcc ctctagactc gagcggccgc cactgtgctg
     2881 gatatctgca gaattccacc acactggact agtggatccg ggccgcatgg tgagcaaggg
     2941 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3001 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3061 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3121 gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3181 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3241 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3301 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3361 ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     3421 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3481 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3541 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3601 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggta ccggaggcgc
     3661 cgctgatact cgcattggtg taacaatcta taagtacgac gataacgcga tgtctgtagt
     3721 gcgcaaggct attgagcaag atgcgaaagc cgcgccagat gttcagctgc tgatgaatga
     3781 ttctcagaat gaccagtcca agcagaacga tcagatcgac gtattgctgg cgaaaggggt
     3841 gaaggcactg gcaatcaacc tggttgaccc ggcagctgcg ggtacggtga ttgagaaagc
     3901 gcgtgggcaa aacgtgccgg tggttttctt caacaaagaa ccgtctcgta aggcgctgga
     3961 tagctacgac aaagcctact acgttggcac tgactccaaa gagtccggca ttattcaagg
     4021 cgatttgatt gctaaacact gggcggcgaa tcagggttgg gatctgaaca aagacggtca
     4081 gattcagttc gtactgctga aaggtgaacc gggccatccg gatgcagaag cacgtaccac
     4141 ttacgtgatt aaagaattga acgataaagg catcaaaact gaacagttac agttagatac
     4201 cgcaatgtgg gacaccgctc aggcgaaaga taagatggac gcctggctgt ctggcccgaa
     4261 cgccaacaaa atcgaagtgg ttatcgccaa caacgatgcg atggcaatgg gcgcggttga
     4321 agcgctgaaa gcacacaaca agtccagcat tccggtgttt ggcgtcgatg cgctgccaga
     4381 agcgctggcg ctggtgaaat ccggtgcact ggcgggcacc gtactgaacg atgctaacaa
     4441 ccaggcgaaa gcgacctttg atctggcgaa aaacctggcc gatggtaaag gtgcggctga
     4501 tggcaccaac tggaaaatcg acaacaaagt ggtccgcgta ccttatgttg gcgtagataa
     4561 agacaacctg gctgaattca gcaagaaagg cgccggtacc ggtggaatgg tgagcaaggg
     4621 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4681 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4741 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     4801 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     4861 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4921 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4981 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5041 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     5101 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     5161 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     5221 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5281 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaagc ttaagtttaa
     5341 accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5521 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     5581 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     5641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     6061 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6121 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6241 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6301 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     6361 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     6421 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6481 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6541 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6601 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     6661 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6721 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6781 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6841 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     6901 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     6961 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     7021 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7081 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7141 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7201 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7261 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     7321 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7381 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     7441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7561 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7621 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7681 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7741 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7801 agaggcggtt tgcgtattgg gcgct
//
