LOCUS       sequence-389433-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of human ATG3 Y209A .
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_Y209A
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        102..481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        482..685
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     636..656
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          821..953
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     936..955
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     997..1016
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        997..1015
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1025..1034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1031..1921
                     /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             1934..1954
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2959..2977)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3009..3130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3020..3039)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3074..3093
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3375..3732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3375..3566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3398..3418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3583..3718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3783..4577
                     /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(3837..3856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4447..4466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4641..4689
                     /note="synthetic polyadenylation signal"
     promoter        4845..4949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4950..5018,5019..5810)
                     /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(5168..5187)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5968..6556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6048..6067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6672..6955
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
       61 gtacatttat attggctcat gtccaatatg accgccatgt tggcattgat tattgactag
      121 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      181 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      241 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      301 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      361 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      421 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      481 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      541 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      601 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      661 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcacta gaagctttat
      721 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc
      781 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
      841 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
      901 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
      961 ctcccagttc aattacagct cttaaggcta gagtattaat acgactcact atagggctag
     1021 caaagccacc atggcagaaa tcggtactgg ctttccattc gacccccatt atgtggaagt
     1081 cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt
     1141 cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc
     1201 gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct
     1261 gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct
     1321 ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa
     1381 gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac
     1441 ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt
     1501 cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt
     1561 cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt
     1621 tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa
     1681 catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct
     1741 gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa
     1801 aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga
     1861 caacccggac ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg
     1921 cgagccaacc actgaggatc tgtactttca gagcgataac gcgatcgctt ccgaattcgg
     1981 atccatgcag aatgtgatta atactgtgaa gggaaaggca ctggaagtgg ctgagtacct
     2041 gaccccggtc ctcaaggaat caaagtttaa ggaaacaggt gtaattaccc cagaagagtt
     2101 tgtggcagct ggagatcacc tagtccacca ctgtccaaca tggcaatggg ctacagggga
     2161 agaattgaaa gtgaaggcat acctaccaac aggcaaacaa tttttggtaa ccaaaaatgt
     2221 gccgtgctat aagcggtgca aacagatgga atattcagat gaattggaag ctatcattga
     2281 agaagatgat ggtgatggcg gatgggtaga tacatatcac aacacaggta ttacaggaat
     2341 aacggaagcc gttaaagaga tcacactgga aaataaggac aatataaggc ttcaagattg
     2401 ctcagcacta tgtgaagagg aagaagatga agatgaagga gaagctgcag atatggaaga
     2461 atatgaagag agtggattgt tggaaacaga tgaggctacc ctagatacaa ggaaaatagt
     2521 agaagcttgt aaagccaaaa ctgatgctgg cggtgaagat gctattttgc aaaccagaac
     2581 ttatgacctt tacatcactt atgataaagc gtaccagact ccacgattat ggttgtttgg
     2641 ctatgatgag caacggcagc ctttaacagt tgagcacatg tatgaagaca tcagtcagga
     2701 tcatgtgaag aaaacagtga ccattgaaaa tcaccctcat ctgccaccac ctcccatgtg
     2761 ttcagttcac ccatgcaggc atgctgaggt gatgaagaaa atcattgaga ctgttgcaga
     2821 aggaggggga gaacttggag ttcatatgta tcttcttatt ttcttgaaat ttgtacaagc
     2881 tgtcattcca acaatagaat atgactacac aagacacttc acaatgtaag cggccgcttg
     2941 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggc cgcttcgagc
     3001 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3061 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3121 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3181 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     3241 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3301 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3361 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3541 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3601 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3661 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3721 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     3781 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3841 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3901 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3961 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4021 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4081 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4141 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4201 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4261 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4321 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4381 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4441 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4501 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4561 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4621 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4681 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     4741 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     4801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     5161 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5341 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     5401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5461 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     5821 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6541 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6661 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     6721 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     6781 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     6841 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     6901 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     6961 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatcttc aatattggcc
     7021 attagccata ttattcattg gtta
//
