LOCUS       sequence-351798-        5757 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 5757)
  AUTHORS   Rhee JM, Pirity MK, Lackan CS, Long JZ, Kondoh G, Takeda J, 
            Hadjantonakis AK
  TITLE     In vivo imaging and differential localization of lipid-modified 
            GFP-variant fusions in embryonic stem cells and mice.
  JOURNAL   Genesis. 2006 Apr;44(4):202-18.
  PUBMED    16604528
REFERENCE   2  (bases 1 to 5757)
  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: "Genesis."; 
            date: "2006-04"; volume: "44(4)"; pages: "202-18"
COMMENT     Sequence Label: pCAG:GPI-GFP
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..650
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          651..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1675..1694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1869..1878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1875..1877,1878..1880,1881..2588)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1920..1941)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2181..2200)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2528..2549
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2784..2803)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2849..2904
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2903..2922)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3265..3281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3265..3281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3278..3300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3289..3305
                     /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(3313..3319,3320..3337,3338..3343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3358..3379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3438..3633
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3484..3619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3639..3773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3676..3695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3730..3749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3841..3858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4012..4600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4092..4111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4771..5562,5563..5631))
                     /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     5394..5413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5632..5736)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
      421 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
      481 gtgcagcgat gggggcgggg gggggggggg gggggcgggg cgaggggcgg ggcggggcga
      541 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      601 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      661 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      721 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      781 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      841 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      901 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      961 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1021 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1141 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1201 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1261 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1441 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1501 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttat tgtgctgtct catcattttg gcaaagaagt accgagctcg gatccactag
     1741 taacggccgc cagtgtgctg gaattcggct tttagggcag gagtatggta gagatgctgc
     1801 caactgtcgc tgtgctggtc ttggcagtgt ccgtggttgc caaggataac accacgctgc
     1861 aggaattcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1921 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1981 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2041 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2101 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2161 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2221 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2281 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2341 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2401 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2461 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2521 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggatg
     2581 aactatacca actcgagaat ggcggcataa gcctgctggt tcagaacaca tcctggatgc
     2641 tgctgctgct gctttccctc tccctcctcc aagccctgga cttcatttct ctgtaagcgg
     2701 caattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     2761 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     2821 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     2881 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     2941 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     3001 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     3061 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     3121 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     3181 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     3241 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3421 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3541 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3601 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     3661 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3721 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     3781 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4441 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgcc
//
