LOCUS       sequence-224747-        5364 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 5364)
  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 5364)
  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: pmCherry-C1-RanQ69L
FEATURES             Location/Qualifiers
     source          1..5364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      602..611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             611..634
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             635..1339
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(777..795)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1045..1064
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2159..2280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2196..2215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2250..2269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2287..2742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2424..2445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2636..2655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2769..2873
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2875..3232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2875..3066
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2898..2918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3083..3218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3267..4061
                     /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             3267..4061
                     /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(3321..3340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3931..3950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4249..4268)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4293..4340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4669..5257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5158..5177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agccaccatg gactataagg acgatgatga caaggtgagc aagggcgagg aggataacat
      661 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      721 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
      781 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      841 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
      901 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
      961 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1021 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1081 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1141 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1201 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1261 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1321 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attctggaat
     1381 ggctgcgcag ggagagcccc aggtccagtt caaacttgta ttggttggtg atggtggtac
     1441 tggaaaaacg accttcgtga aacgtcattt gactggtgaa tttgagaaga agtatgtagc
     1501 caccttgggt gttgaggttc atcccctagt gttccacacc aacagaggac ctattaagtt
     1561 caatgtatgg gacacagccg gcctggagaa attcggtgga ctgagagatg gctattatat
     1621 ccaagcccag tgtgccatca taatgtttga tgtaacatcg agagttactt acaagaatgt
     1681 gcctaactgg catagagatc tggtacgagt gtgtgaaaac atccccattg tgttgtgtgg
     1741 caacaaagtg gatattaagg acaggaaagt gaaggcgaaa tccattgtct tccaccgaaa
     1801 gaagaatctt cagtactacg acatttctgc caaaagtaac tacaactttg aaaagccctt
     1861 cctctggctt gctaggaagc tcattggaga ccctaacttg gaatttgttg ccatgcctgc
     1921 tctcgcccca ccagaagttg tcatggaccc agctttggca gcacagtatg agcacgactt
     1981 agaggttgct cagacaactg ctctcccgga tgaggatgat gacctgtgag gatccaccgg
     2041 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2101 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2161 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2221 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2281 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2341 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2401 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2461 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2521 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2581 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2641 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2701 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2881 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3061 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3121 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3181 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3241 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3301 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3361 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3421 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3481 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3541 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3601 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3661 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3721 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3781 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3841 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3901 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3961 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4021 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4081 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4201 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4261 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4321 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4381 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4441 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4501 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5281 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5341 ttctgtggat aaccgtatta ccgc
//
