LOCUS       sequence-200861-        6412 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 6412)
  AUTHORS   Kazgan N, Williams T, Forsberg LJ, Brenman JE
  TITLE     Identification of a nuclear export signal in the catalytic subunit 
            of AMP-activated protein kinase.
  JOURNAL   Mol Biol Cell. 2010 Oct 1. 21(19):3433-42.
  PUBMED    20685962
REFERENCE   2  (bases 1 to 6412)
  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: "Mol Biol 
            Cell. 2010 Oct 1. 21(19):3433-42."
COMMENT     Sequence Label: pEGFP-C1-PRKAA2
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        141..444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        445..648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     599..619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(693..695,696..698,699..1409)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(693..695,696..698,699..1409)
                     /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(738..759)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(999..1018)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1346..1367
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3280..3401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3317..3336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3371..3390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3408..3863)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3545..3566)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3757..3776
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3890..3994
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3996..4353
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3996..4187
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4019..4039)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4204..4339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4388..5182
                     /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             4388..5182
                     /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(4442..4461)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5052..5071
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5370..5389)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5414..5461
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5790..6378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6279..6298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
       61 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
      121 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      181 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      241 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      301 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      361 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      421 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      481 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      541 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      601 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      661 cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
      721 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      781 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      841 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      901 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      961 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1021 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1081 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1141 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1201 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1261 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1321 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1381 tcactctcgg catggacgag ctgtacaagt ccggccggac tcagatctcg agctcaagct
     1441 tcgaattcaa catggctgag aagcagaagc acgacgggcg tgtgaagatc ggacactacg
     1501 tgctggggga caccctgggc gtcggcacct tcggcaaagt gaagattgga gaacatcaat
     1561 tgacaggcca taaagtggca gttaagatct taaatagaca gaagattcgc agtttagatg
     1621 ttgttggaaa aataaaacga gaaattcaaa atcttaaact ctttcgtcat cctcatatta
     1681 tcaaactcta ccaagtgatc agcactccaa cagacttttt tatggtaatg gaatatgtgt
     1741 ctggaggtga attgttcgac tacatctgta aacacgggag ggttgaagag gtggaagctc
     1801 gccggctctt ccagcagatt ctgtctgccg tggactactg tcacaggcac atggttgtcc
     1861 acagggacct gaagccagag aacgtgttgc tggacgccca gatgaatgct aagatagctg
     1921 acttcggact ctctaatatg atgtcagatg gtgaatttct acgaactagc tgtggatcgc
     1981 caaattatgc agcaccggag gtcatctcag gaaggctgta tgcgggtcct gaggttgata
     2041 tctggagctg tggtgttatc ctgtatgccc ttctctgtgg caccctcccg ttcgacgatg
     2101 agcacgtgcc tacgctcttt aagaagatcc gagggggtgt gttctacatc ccggagtatc
     2161 tcaaccgttc tattgccact ctgctgatgc acatgctgca ggtggacccc ttgaagcgag
     2221 caactatcaa agacatacga gagcatgaat ggtttaaaca ggatttgccc agttacctct
     2281 ttcctgaaga cccctcctat gatgctaacg tcattgatga tgaggctgtg aaagaagtat
     2341 gtgaaaaatt tgagtgtaca gaatcagaag tgatgaacag tttatacagt ggtgaccctc
     2401 aagaccagct cgcagtggct tatcatctca tcattgacaa tcggagaata atgaaccaag
     2461 ccagtgagtt ctacctcgcc tccagtcctc caacgggttc cttcatggac gatagtgcca
     2521 tgcatattcc ccccggcctg aaaccacatc ctgaaaggat gccacctctc atagcagaca
     2581 gccccaaagc acgctgtcca ctggatgcac tcaacacaac taagcccaaa tctttagctg
     2641 tgaaaaaagc caagtggcac cttgggatcc gaagccagag caaaccatac gacattatgg
     2701 cggaggtgta ccgagctatg aagcagctgg actttgaatg gaaggtagtg aatgcatacc
     2761 atcttcgagt aagaagaaaa aacccagtga ctggcaatta cgtgaaaatg agcttacagc
     2821 tttacctggt tgacaatcgg agctatcttc tagactttaa aagcatcgat gatgaggtgg
     2881 tggagcagag gtctggttct tcaacacctc agcgctcctg ttctgctgcc ggcctccaca
     2941 gacctcggtc aagtgtcgat tccagcacag ccgagaacca ttcactgtct ggctctctca
     3001 ctggttcttt gactggcagc actttgtcct ccgcttcccc gcgcctgggc agtcatacca
     3061 tggatttttt tgaaatgtgc gccagtctta tcactgcttt agcccgttga ggatccataa
     3121 agggcgaatt ccacattggg ctgcagcccg ggatccaccg gatctagata actgatcata
     3181 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3241 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3301 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3361 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     3421 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3481 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3541 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3601 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3661 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3721 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3781 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3841 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3901 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3961 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4021 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4141 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4201 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4261 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4321 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4381 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4441 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4501 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4561 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4621 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4681 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4741 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4801 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4861 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4921 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4981 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5041 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5101 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5161 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5221 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5281 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5341 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5401 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5461 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5521 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5581 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct gg
//
