LOCUS       sequence-403836-        6177 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 6177)
  AUTHORS   Watton SJ, Downward J
  TITLE     Akt/PKB localisation and 3' phosphoinositide generation at sites of 
            epithelial cell-matrix and cell-cell interaction.
  JOURNAL   Curr Biol. 1999 Apr 22;9(8):433-6.
  PUBMED    10226029
REFERENCE   2  (bases 1 to 6177)
  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: "Curr 
            Biol."; date: "1999-04-22"; volume: "9(8)"; pages: "433-6"
COMMENT     Sequence Label: pEGFP-Akt-DD
FEATURES             Location/Qualifiers
     source          1..6177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      721..730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(727..729,730..732,733..1443)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(727..729,730..732,733..1443)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(772..793)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1033..1052)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1380..1401
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1471..1497
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3079..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3170..3189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3207..3662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3344..3365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3556..3575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3689..3793
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3795..4152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3795..3986
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3818..3838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4003..4138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4187..4981
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4187..4981
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4241..4260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4851..4870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5169..5188)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5213..5260
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      721 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      781 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      841 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      901 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      961 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1021 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1081 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1141 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1201 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1261 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1321 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1381 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1441 aagggtggag gtggaggagg cgtcgacatg tacccatacg atgttccaga ttacgcttct
     1501 agaatgaacg acgtagccat tgtgaaggag ggctggctgc acaaacgagg ggaatatatt
     1561 aaaacctggc ggccacgcta cttcctcctc aagaacgatg gcacctttat tggctacaag
     1621 gaacggcctc aggatgtgga tcagcgagag tccccactca acaacttctc agtggcacaa
     1681 tgccagctga tgaagacaga gcggccaagg cccaacacct ttatcatccg ctgcctgcag
     1741 tggaccacag tcattgagcg caccttccat gtggaaacgc ctgaggagcg ggaagaatgg
     1801 gccaccgcca ttcagactgt ggccgatgga ctcaagaggc aggaagaaga gacgatggac
     1861 ttccgatcag gctcacccag tgacaactca ggggctgaag agatggaggt gtccctggcc
     1921 aagcccaagc accgtgtgac catgaacgag tttgagtacc tgaaactact gggcaagggc
     1981 acctttggga aagtgattct ggtgaaagag aaggccacag gccgctacta tgccatgaag
     2041 atcctcaaga aggaggtcat cgtcgccaag gatgaggttg cccacacgct tactgagaac
     2101 cgtgtcctgc agaactctag gcatcccttc cttacggccc tcaagtactc attccagacc
     2161 cacgaccgcc tctgctttgt catggagtat gccaacgggg gcgagctctt cttccacctg
     2221 tctcgagagc gcgtgttctc cgaggaccgg gcccgcttct atggtgcgga gattgtgtct
     2281 gccctggact acttgcactc cgagaagaac gtggtgtacc gggacctgaa gctggagaac
     2341 ctcatgctgg acaaggacgg gcacatcaag ataacggact tcgggctgtg caaggagggg
     2401 atcaaggatg gtgccactat gaaggatttc tgcggaacgc cggagtacct ggcccctgag
     2461 gtgctggagg acaacgacta cggccgtgca gtggactggt gggggctggg cgtggtcatg
     2521 tatgagatga tgtgtggccg cctgcccttc tacaaccagg accacgagaa gctgttcgag
     2581 ctgatcctca tggaggagat ccgcttcccg cgcacactcg gccctgaggc caagtccctg
     2641 ctctccgggc tgctcaagaa ggaccctaca cagaggctcg gtgggggctc tgaggatgcc
     2701 aaggagatca tgcagcaccg gttctttgcc aacatcgtgt ggcaggatgt gtatgagaag
     2761 aagctgagcc cacctttcaa gccccaggtc acctctgaga ctgacaccag gtatttcgat
     2821 gaggagttca cagctcagat gatcaccatc acgccgcctg atcaagatga cagcatggag
     2881 tgtgtggaca gtgagcggag gccgcacttc ccccagttcg actactcagc cagtggcaca
     2941 gcctgaggcg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     3001 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3061 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3181 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3241 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3301 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3361 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3421 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3481 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3541 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3601 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3661 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3721 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3781 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3961 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4021 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4081 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4141 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5161 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5221 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5281 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5341 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5401 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaa
//
