LOCUS       sequence-217935-        9921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient/stable overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9921)
  TITLE     Proszynski unpublished
REFERENCE   2  (bases 1 to 9921)
  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"
COMMENT     Sequence Label: FFTTH-Amotl2
FEATURES             Location/Qualifiers
     source          1..9921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        90..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             646..669
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             679..702
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             712..732
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             742..762
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             772..798
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3259..3380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3350..3369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6571..6673
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6762..6779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6933..7521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7013..7032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             8353..9147
                     /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             8353..9147
                     /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(8407..8426)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9017..9036
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(9195..9575)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9257..9278)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9469..9488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   9613..9715
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     join(9803..9921,1..32)
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 tgaataattt tgtgttactc atagcgcgta atactgtaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      601 ttagtgaacc gtcagatccg ctagagatct ggtaccgtcg acatggatta caaggatgac
      661 gacgataagt caggtggaga ttacaaggat gacgacgata agagcggccg cgaaaatctg
      721 tactttcaag gcgcatcagg agaaaatctg tactttcaag gcggatctgg atacccctac
      781 gacgtccccg actacgccaa gcttatgagg acactggaag actcctcagg gacagtcctg
      841 caccgtctca tccaagagca gctgcgctac ggcaacctga cagagactcg cactctgctg
      901 gctatccagc agcaggccct gcggggtggg gctggagctg ggggcacggg gagcccccag
      961 gcttccttgg agatcggagc acccgaggac agtcaggtgc tgcagcaagc caccaggcag
     1021 gggccccagg gccaggagca tcagggtgga gagacccacc tggcagagaa caggctgtac
     1081 cggctgtgcc cacagcccag caaaggagaa gagttgccca cctatgagga ggccaaagcc
     1141 cattcgcagt actacgcagc gcagcaggca gggtcccggc cgcatgttgg ggaccgggat
     1201 cctagaggag gggtgtccgg aggcggccgg cgacaggatg aagctcttcg agagctgagg
     1261 catggccatg tgcgctcctt gagtgaacgg cttctgcaac tgtccctgga aagaaacggt
     1321 gctcgggtcc ccagccacat gagctcttcc cacagcttcc ctcagctggc ccgcagccag
     1381 cagggccccc aaccccgagg gcccccagct gagggcccag agccccgcgg gccaccacct
     1441 cagtacccac acgctgtaat ggctcaggag actgcggctg tcactgaccc aagataccga
     1501 ccccgaagca gcccacactt ccagcatgcc gaagtcagga tcctgcaggc ccaggtacca
     1561 ccggtgttcc tccagcagca gcagtaccag tacctgccgc agccccagga gcactctcca
     1621 cccctccacc cggcagctct gggccatgga cccccaagct cctttggtcc acctgcagtg
     1681 gagggaccac ccagtgccca ggccaccttg ggcagtgccc acctggccca gatggagact
     1741 gtactgaggg agaatgccag gctgcagagg gacaatgagc gattgcagag agagctggag
     1801 agcacttcag agaaggctgg ccgcatagaa aagctggaaa atgaaatcca gcggctctct
     1861 gaggcccacg agagcctgat gaggacctct tccaagcgtg aggccctgga gaagaccatg
     1921 aggaacaaga tggacggtga gatgagacgg ttgcaggact tcaaccgaga ccttagagag
     1981 agattggaat cggcaaaccg ccacctggca agcaagaccc aggaagccca ggcgggcagt
     2041 caggacatgg tggcgaaact gcttgcccag agctatgagc agcaacagga acaggagaag
     2101 ctggagcggg agatggcact gctgcgtggt gccatcgagg accagcggcg acatgctgaa
     2161 ctgctggagc aggctctggg caatgcacaa agccgtgccg cccgggctga agaggagcta
     2221 cgcaaaaagc aggcctatgt ggagaaggtg gagcggctgc agcaggcact ggggcagttg
     2281 caggctgcct gtgaaaagcg agagcagttg gagctgcgtc tgcgcacgcg cctggagcag
     2341 gaactcaaag ccttgcgtgc acagcagagg cagacaggca ccctcgcagg tggtggcggc
     2401 agccatggtg ggtccgccga gctcagtgcc ctgcggctgt ctgaacagct gcgggagaag
     2461 gaggaacaga tcctggctct agaggcggac atgaccaagt gggaacagaa gtatttggaa
     2521 gaacgggcta tgaggcagtt cgccatggac gcggctgcca ctgcggctgc ccagcgcgat
     2581 accactctca tccggcactc cccccagccc tcgcccagca gcagtttcaa cgagggcctg
     2641 ctgccaggca accacaggca ccaggagatg gagagcagat tgaaggtgct ccatgctcag
     2701 atcctagaga aggatgcggt gatcaaggtc cttcagcagc gctccaggaa agaccctggc
     2761 aaggccaccc agggcaccct acggcctgcc aagtcggtgc cgtccatctt cgcggctgca
     2821 gtgggaactc agggctggca agggctcgta tccagtgagc ggcaaactga tgcacggcca
     2881 gcgggagacc gggtcccagc agaggagcct ccggccacag ctcctctccc tgcccacacc
     2941 aaacatggca gcagagacgg gagcacccag acggatggcc ctgcagacaa cacctctgcc
     3001 tgcttggcct cagaacccga tggcctcctg gggtgcaaca gtagccagag gacaccctct
     3061 ctggactcta tagctgcaac cagagtccag gatctgtcag acatggtaga aatactgatc
     3121 tgagatatcc gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     3181 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3241 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3361 aaactcatca atgtatctta acgcggatct gggcgtggtt aagggtggga aagaatatat
     3421 aaggtggggg tcttatgtag ttttgtatct gttttgcagc agccgccgcc gccatgagca
     3481 ccaactcgtt tgatggaagc attgtgagct catatttgac aacgcgcatg cccccatggg
     3541 ccggggtgcg tcagaatgtg atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa
     3601 actctactac cttgacctac gagaccgtgt ctggaacgcc gttggagact gcagcctccg
     3661 ccgccgcttc agccgctgca gccaccgccc gcgggattgt gactgacttt gctttcctga
     3721 gcccgcttgc aagcagtgca gcttcccgtt catccgcccg cgatgacaag ttgacggctc
     3781 ttttggcaca attggattct ttgacccggg aacttaatgt cgtttctcag cagctgttgg
     3841 atctgcgcca gcaggtttct gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca
     3901 taaataaaaa accagactct gtttggattt ggatcaagca agtgtcttgc tgtctttatt
     3961 taggggtttt gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg agggtcctgt
     4021 gtattttttc caggacgtgg taaaggtgac tctggatgtt cagatacatg ggcataagcc
     4081 cgtctctggg gtggaggtag caccactgca gagcttcatg ctgcggggtg gtgttgtaga
     4141 tgatccagtc gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc
     4201 tgattgccag gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt
     4261 gcatacgtgg ggatatgaga tgcatcttgg actgtatttt taggttggct atgttcccag
     4321 ccatatccct ccggggattc atgttgtgca gaaccaccag cacagtgtat ccggtgcact
     4381 tgggaaattt gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag acgcccttgt
     4441 gacctccaag attttccatg cattcgtcca taatgatggc aatgggccca cgggcggcgg
     4501 cctgggcgaa gatatttctg ggatcactaa cgtcatagtt gtgttccagg atgagatcgt
     4561 cataggccat ttttacaaag cgcgggcgga gggtgccaga ctgcggtata atggttccat
     4621 ccggcccagg ggcgtagtta ccctcacaga tttgcatttc ccacgctttg agttcagatg
     4681 gggggatcat gtctacctgc ggggcgatga agaaaacggt ttccggggta ggggagatca
     4741 gctgggaaga aagcaggttc ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa
     4801 tcacacctat taccggctgc aactggtagt taagagagct gcagctgccg tcatccctga
     4861 gcaggggggc cacttcgtta agcatgtccc tgactcgcat gttttccctg accaaatccg
     4921 ccagaaggcg ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag tttttcaacg
     4981 gtttgagacc gtccgccgta ggcatgcttt tgagcgtttg accaagcagt tccaggcggt
     5041 cccacagctc ggtcacctgc tctacggcat ctcgatccag catatctcct cgtttcgcgg
     5101 gttggggcgg ctttcgctgt acggcagtag tcggtgctcg tccagacggg ccagggtcat
     5161 gtctttccac gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc
     5221 tccgggctgc gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg
     5281 ccggtcttcg ccctgcgcgt cggccaggta gcatttgacc atggtgtcat agtccagccc
     5341 ctccgcggcg tggcccttgg cgcgcagctt gcccttggag gaggcgccgc acgaggggca
     5401 gtgcagactt ttgagggcgt agagcttggg cgcgagaaat accgattccg gggagtaggc
     5461 atccgcgccg caggccccgc agacggtctc gcattccacg agccaggtga gctctggccg
     5521 ttcggggtca aaaaccaggt ttcccccatg ctttttgatg cgtttcttac ctctggtttc
     5581 catgagccgg tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt atacagactt
     5641 gagagggagt ttaaacgaat tcaatagctt gttgcatggg cggcgatata aaatgcaagg
     5701 tgctgctcaa aaaatcaggc aaagcctcgc gcaaaaaaga aagcacatcg tagtcatgct
     5761 catgcagata aaggcaggta agctccggaa ccaccacaga aaaagacacc atttttctct
     5821 caaacatgtc tgcgggtttc tgcataaaca caaaataaaa taacaaaaaa acatttaaac
     5881 attagaagcc tgtcttacaa caggaaaaac aacccttata agcataagac ggactacggc
     5941 catgccggcg tgaccgtaaa aaaactggtc accgtgatta aaaagcacca ccgacagctc
     6001 ctcggtcatg tccggagtca taatgtaaga ctcggtaaac acatcaggtt gattcacatc
     6061 ggtcagtgct aaaaagcgac cgaaatagcc cgggggaata catacccgca ggcgtagaga
     6121 caacattaca gcccccatag gaggtataac aaaattaata ggagagaaaa acacataaac
     6181 acctgaaaaa ccctcctgcc taggcaaaat agcaccctcc cgctccagaa caacatacag
     6241 cgcttccaca gcggcagcca taacagtcag ccttaccagt aaaaaagaaa acctattaaa
     6301 aaaacaccac tcgacacggc accagctcaa tcagtcacag tgtaaaaaag ggccaagtgc
     6361 agagcgagta tatataggac taaaaaatga cgtaacggtt aaagtccaca aaaaacaccc
     6421 agaaaaccgc acgcgaacct acgcccagaa acgaaagcca aaaaacccac aacttcctca
     6481 aatcgtcact tccgttttcc cacgttacgt cacttcccat tttaagaaaa ctacaattcc
     6541 caacacatac aagttactcc gccctaaaac ctacgtcacc cgccccgttc ccacgccccg
     6601 cgccacgtca caaactccac cccctcatta tcatattggc ttcaatccaa aataaggtat
     6661 attattgatg atgttaatta acatgcatgg atccatatgc ggtgtgaaat accgcacaga
     6721 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     6781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7981 agtttgcgca acgttgttgc cattgctgca gccatgagat tatcaaaaag gatcttcacc
     8041 tagatccttt tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc
     8101 agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc
     8161 agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa
     8221 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
     8281 ttcttgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga
     8341 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     8401 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     8461 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     8521 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     8581 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     8641 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     8701 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     8761 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     8821 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     8881 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     8941 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     9001 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     9061 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     9121 tatcgccttc ttgacgagtt cttctgaatt ttgttaaaat ttttgttaaa tcagctcatt
     9181 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     9241 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     9301 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     9361 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     9421 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     9481 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     9541 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gatcgaatta
     9601 attcttaatt aacatcatca ataatatacc ttattttgga ttgaagccaa tatgataatg
     9661 agggggtgga gtttgtgacg tggcgcgggg cgtgggaacg gggcgggtga cgtagtagtg
     9721 tggcggaagt gtgatgttgc aagtgtggcg gaacacatgt aagcgacgga tgtggcaaaa
     9781 gtgacgtttt tggtgtgcgc cggtgtacac aggaagtgac aattttcgcg cggttttagg
     9841 cggatgttgt agtaaatttg ggcgtaaccg agtaagattt ggccattttc gcgggaaaac
     9901 tgaataagag gaagtgaaat c
//
