LOCUS       sequence-389126-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of human ATG3 T244A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  TITLE     Hurley ATG3 plasmids
REFERENCE   2  (bases 1 to 7044)
  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: pHTN-Halo-TEV-ATG3_T244A
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        451..654
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     605..625
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          790..922
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     905..924
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     966..985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        966..984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..1890
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1903..1923
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2928..2946)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2978..3099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2989..3008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3043..3062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3344..3701
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3344..3535
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3367..3387)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3552..3687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3752..4546
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3806..3825)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4416..4435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4610..4658
                     /note="synthetic polyadenylation signal"
     promoter        4814..4918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4919..4987,4988..5779)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5137..5156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5937..6525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6017..6036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6641..6924
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
       61 ccgccatgtt ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      121 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      181 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      241 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      301 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      361 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
      421 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      481 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      541 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      601 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      661 gtgaaccgtc agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac
      721 gcagtcagtg cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc
      781 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
      841 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
      901 cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag
      961 agtattaata cgactcacta tagggctagc aaagccacca tggcagaaat cggtactggc
     1021 tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt
     1081 ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg
     1141 tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc
     1201 ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc
     1261 atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg
     1321 ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca
     1381 tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag
     1441 accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
     1501 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
     1561 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
     1621 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
     1681 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
     1741 ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc
     1801 ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg
     1861 cgctggctgt cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag
     1921 agcgataacg cgatcgcttc cgaattcgga tccatgcaga atgtgattaa tactgtgaag
     1981 ggaaaggcac tggaagtggc tgagtacctg accccggtcc tcaaggaatc aaagtttaag
     2041 gaaacaggtg taattacccc agaagagttt gtggcagctg gagatcacct agtccaccac
     2101 tgtccaacat ggcaatgggc tacaggggaa gaattgaaag tgaaggcata cctaccaaca
     2161 ggcaaacaat ttttggtaac caaaaatgtg ccgtgctata agcggtgcaa acagatggaa
     2221 tattcagatg aattggaagc tatcattgaa gaagatgatg gtgatggcgg atgggtagat
     2281 acatatcaca acacaggtat tacaggaata acggaagccg ttaaagagat cacactggaa
     2341 aataaggaca atataaggct tcaagattgc tcagcactat gtgaagagga agaagatgaa
     2401 gatgaaggag aagctgcaga tatggaagaa tatgaagaga gtggattgtt ggaaacagat
     2461 gaggctaccc tagatacaag gaaaatagta gaagcttgta aagccaaaac tgatgctggc
     2521 ggtgaagatg ctattttgca aaccagaact tatgaccttt acatcactta tgataaatat
     2581 taccagactc cacgattatg gttgtttggc tatgatgagc aacggcagcc tttaacagtt
     2641 gagcacatgt atgaagacat cagtcaggat catgtgaaga aagcggtgac cattgaaaat
     2701 caccctcatc tgccaccacc tcccatgtgt tcagttcacc catgcaggca tgctgaggtg
     2761 atgaagaaaa tcattgagac tgttgcagaa ggagggggag aacttggagt tcatatgtat
     2821 cttcttattt tcttgaaatt tgtacaagct gtcattccaa caatagaata tgactacaca
     2881 agacacttca caatgtaagc ggccgcttgc tgagttggct gctgccaccg ctgagcaata
     2941 actagcataa ccccttggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     3001 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3061 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3121 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     3181 aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     3241 tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat
     3301 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
     3361 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3481 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3541 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3601 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3661 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca
     3721 caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg
     3781 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3841 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3901 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3961 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4021 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4081 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4141 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4201 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4261 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4321 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4381 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4441 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4501 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4561 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc
     4621 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct
     4681 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     4741 gcaccgtttc tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt
     4801 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4921 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4981 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5041 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5101 agaacgtttt ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg
     5161 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5221 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5281 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg
     5341 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5401 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5461 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5521 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5581 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5641 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5701 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5761 actgattaag cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct
     5821 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6361 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     6541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6601 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac
     6661 gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa
     6721 gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat
     6781 caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg
     6841 tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt
     6901 actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc
     6961 gtagaggatc gagatcttca atattggcca ttagccatat tattcattgg ttatatagca
     7021 taaatcaata ttggctattg gcca
//
