LOCUS       sequence-389409-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of human ATG3 Y210A .
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_Y210A
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..487
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        488..691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     642..662
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          827..959
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     942..961
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1003..1022
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1003..1021
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1031..1040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1037..1927
                     /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             1940..1960
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2965..2983)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3015..3136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3026..3045)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3080..3099
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3381..3738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3381..3572
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3404..3424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3589..3724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3789..4583
                     /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(3843..3862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4453..4472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4647..4695
                     /note="synthetic polyadenylation signal"
     promoter        4851..4955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4956..5024,5025..5816)
                     /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(5174..5193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5974..6562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6054..6073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6678..6961
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
       61 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttggc attgattatt
      121 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      181 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      241 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      301 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      361 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      421 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      481 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
      541 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      601 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      661 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcactagaag
      721 ctttattgcg gtagtttatc acagttaaat tgctaacgca gtcagtgctt ctgacacaac
      781 agtctcgaac ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt
      841 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
      901 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
      961 tgtccactcc cagttcaatt acagctctta aggctagagt attaatacga ctcactatag
     1021 ggctagcaaa gccaccatgg cagaaatcgg tactggcttt ccattcgacc cccattatgt
     1081 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     1141 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     1201 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     1261 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     1321 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     1381 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     1441 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     1501 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     1561 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     1621 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     1681 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     1741 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     1801 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     1861 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat
     1921 ttccggcgag ccaaccactg aggatctgta ctttcagagc gataacgcga tcgcttccga
     1981 attcggatcc atgcagaatg tgattaatac tgtgaaggga aaggcactgg aagtggctga
     2041 gtacctgacc ccggtcctca aggaatcaaa gtttaaggaa acaggtgtaa ttaccccaga
     2101 agagtttgtg gcagctggag atcacctagt ccaccactgt ccaacatggc aatgggctac
     2161 aggggaagaa ttgaaagtga aggcatacct accaacaggc aaacaatttt tggtaaccaa
     2221 aaatgtgccg tgctataagc ggtgcaaaca gatggaatat tcagatgaat tggaagctat
     2281 cattgaagaa gatgatggtg atggcggatg ggtagataca tatcacaaca caggtattac
     2341 aggaataacg gaagccgtta aagagatcac actggaaaat aaggacaata taaggcttca
     2401 agattgctca gcactatgtg aagaggaaga agatgaagat gaaggagaag ctgcagatat
     2461 ggaagaatat gaagagagtg gattgttgga aacagatgag gctaccctag atacaaggaa
     2521 aatagtagaa gcttgtaaag ccaaaactga tgctggcggt gaagatgcta ttttgcaaac
     2581 cagaacttat gacctttaca tcacttatga taaatatgcg cagactccac gattatggtt
     2641 gtttggctat gatgagcaac ggcagccttt aacagttgag cacatgtatg aagacatcag
     2701 tcaggatcat gtgaagaaaa cagtgaccat tgaaaatcac cctcatctgc caccacctcc
     2761 catgtgttca gttcacccat gcaggcatgc tgaggtgatg aagaaaatca ttgagactgt
     2821 tgcagaagga gggggagaac ttggagttca tatgtatctt cttattttct tgaaatttgt
     2881 acaagctgtc attccaacaa tagaatatga ctacacaaga cacttcacaa tgtaagcggc
     2941 cgcttgctga gttggctgct gccaccgctg agcaataact agcataaccc cttggccgct
     3001 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3061 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3121 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3181 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     3241 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     3301 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     3361 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3481 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3661 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3721 ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta
     3781 gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3961 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4561 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4621 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     4681 ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg
     4741 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     4801 tctacgtaat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4861 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4921 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4981 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5041 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5101 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5161 ctttcaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5221 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5281 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5341 ataacactgc ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt
     5401 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5461 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5521 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5581 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5641 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5701 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5761 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg
     5821 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     5881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6541 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6661 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     6721 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     6781 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     6841 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     6901 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     6961 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atcttcaata
     7021 ttggccatta gccatattat tcat
//
