LOCUS       sequence-200863-        6370 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 6370)
  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 6370)
  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-PRKAA1
FEATURES             Location/Qualifiers
     source          1..6370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        464..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     618..638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      706..715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(712..714,715..717,718..1428)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(712..714,715..717,718..1428)
                     /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(757..778)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1018..1037)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1365..1386
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3257..3378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3348..3367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3385..3840)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3522..3543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3734..3753
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3867..3971
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3973..4330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3973..4164
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3996..4016)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4181..4316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4365..5159
                     /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             4365..5159
                     /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(4419..4438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5029..5048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5347..5366)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5391..5438
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5767..6355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6256..6275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
       61 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
      121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      661 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
      721 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      781 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      841 accctgaagt tcatctgcac caccggcaag ctgcccgtcc cctggcccac cctcgtgacc
      901 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      961 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1021 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1081 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1141 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1201 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1261 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1321 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1381 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga
     1441 tctcgagaca tggccgagaa gcagaagcac gacgggcggg tgaagatcgg ccactacatc
     1501 ctgggggaca cgctgggcgt cggcaccttc gggaaagtga aggtgggcaa gcacgagttg
     1561 actggacata aagttgctgt gaagatactc aaccggcaga agattcgaag cctggacgtg
     1621 gtcgggaaaa tccgcagaga gatccagaac ctgaagcttt tcaggcaccc tcatataatc
     1681 aaactgtacc aggtcatcag tacaccgtct gatattttca tggtcatgga atatgtctca
     1741 ggaggagagc tatttgatta tatctgtaaa aatggaaggt tggacgaaaa ggagagtcga
     1801 cgtctgttcc agcagatcct ttctggtgtg gactattgtc acaggcatat ggtggtccac
     1861 agagatttga aacctgaaaa cgtcctgctt gatgcacaca tgaatgcaaa gatagccgac
     1921 ttcggtcttt caaacatgat gtcagatggt gaatttttaa gaacgagctg tggctcgccc
     1981 aattatgctg caccagaagt aatttcagga agattgtacg caggccctga agtagacatc
     2041 tggagcagcg gggtcattct ctatgctttg ctgtgtggaa ctctcccttt tgatgatgac
     2101 cacgtgccaa ctctttttaa gaagatatgt gacgggatat tttatacccc tcagtatttg
     2161 aatccctctg taataagcct tttgaagcat atgctgcagg tagatcctat gaagagggcc
     2221 acaataaaag atatcaggga acatgaatgg tttaagcagg accttccaaa atatctcttt
     2281 cctgaagacc cgtcttatag ttcaaccatg attgatgatg aagccttaaa agaagtgtgt
     2341 gagaagttcg agtgctcaga ggaggaggtc ctcagctgcc tgtacaacag aaaccaccag
     2401 gacccactgg cagttgccta ccacctcata atagacaaca ggagaataat gaacgaagcc
     2461 aaagatttct acttggcaac aagcccaccc gattctttcc tcgatgatca ccatttaact
     2521 cggcctcacc ctgagagagt accattcttg gttgccgaaa caccaagggc ccgacacacc
     2581 ctagatgaat taaacccaca gaaatccaaa caccaaggcg tacggaaggc aaagtggcat
     2641 ttggggattc gaagtcaaag ccgacccaat gacatcatgg cagaagtgtg tagagcaatc
     2701 aagcagttgg actatgaatg gaaggttgta aacccctatt atttgcgtgt gcgaaggaag
     2761 aaccctgtga caagcacatt ttccaaaatg agtctacagc tataccaagt ggatagtagg
     2821 acttacttat tggatttccg aagtattgat gatgagatta cagaagccaa atcagggact
     2881 gctactccac agagatcggg atccatcagc aactatcgat cttgccaaag gagcgactcc
     2941 gacgccgagg ctcaaggaaa gccctcagaa gtctctctta cctcatccgt gacctccctc
     3001 gactcctctc ctgttgacgt agctccaaga ccaggaagtc acacgataga attttttgaa
     3061 atgtgtgcaa atctaattaa aattcttgca cagtaagaat tctgcagtcg acggtaccgc
     3121 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     3181 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3241 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3301 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3361 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3421 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3481 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3541 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3601 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3661 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3721 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3781 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3841 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3961 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4021 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4081 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4141 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4201 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4261 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4321 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4381 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4441 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4501 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4561 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4621 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4681 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4741 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4801 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4861 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4921 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4981 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5041 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5101 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5161 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5221 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5281 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5341 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5401 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5461 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5521 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5581 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5881 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6361 cagcaacgcg
//
