LOCUS       sequence-389430-        7044 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of human ATG3 .
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 Apr 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
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..474
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        475..678
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     629..649
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          814..946
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     929..948
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     990..1009
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        990..1008
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1024..1914
                     /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             1927..1947
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2952..2970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3002..3123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3013..3032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3067..3086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3368..3725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3368..3559
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3391..3411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3576..3711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3776..4570
                     /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(3830..3849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4440..4459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4634..4682
                     /note="synthetic polyadenylation signal"
     promoter        4838..4942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4943..5011,5012..5803)
                     /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(5161..5180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5961..6549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6041..6060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6665..6948
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
       61 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
      121 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      181 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      241 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      301 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
      361 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      421 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      481 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      541 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      601 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
      661 gtctatataa gcagagctgg tttagtgaac cgtcagatca ctagaagctt tattgcggta
      721 gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt ctcgaactta
      781 agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt
      841 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
      901 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag
      961 ttcaattaca gctcttaagg ctagagtatt aatacgactc actatagggc tagcaaagcc
     1021 accatggcag aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc
     1081 gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac
     1141 ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat
     1201 cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat
     1261 ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag
     1321 gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat
     1381 ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac
     1441 gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc
     1501 aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc
     1561 ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc
     1621 gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc
     1681 gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc
     1741 tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg
     1801 cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg
     1861 gacctgatcg gcagcgagat cgcgcgctgg ctgtcgacgc tcgagatttc cggcgagcca
     1921 accactgagg atctgtactt tcagagcgat aacgcgatcg cttccgaatt cggatccatg
     1981 cagaatgtga ttaatactgt gaagggaaag gcactggaag tggctgagta cctgaccccg
     2041 gtcctcaagg aatcaaagtt taaggaaaca ggtgtaatta ccccagaaga gtttgtggca
     2101 gctggagatc acctagtcca ccactgtcca acatggcaat gggctacagg ggaagaattg
     2161 aaagtgaagg catacctacc aacaggcaaa caatttttgg taaccaaaaa tgtgccgtgc
     2221 tataagcggt gcaaacagat ggaatattca gatgaattgg aagctatcat tgaagaagat
     2281 gatggtgatg gcggatgggt agatacatat cacaacacag gtattacagg aataacggaa
     2341 gccgttaaag agatcacact ggaaaataag gacaatataa ggcttcaaga ttgctcagca
     2401 ctatgtgaag aggaagaaga tgaagatgaa ggagaagctg cagatatgga agaatatgaa
     2461 gagagtggat tgttggaaac agatgaggct accctagata caaggaaaat agtagaagct
     2521 tgtaaagcca aaactgatgc tggcggtgaa gatgctattt tgcaaaccag aacttatgac
     2581 ctttacatca cttatgataa atattaccag actccacgat tatggttgtt tggctatgat
     2641 gagcaacggc agcctttaac agttgagcac atgtatgaag acatcagtca ggatcatgtg
     2701 aagaaaacag tgaccattga aaatcaccct catctgccac cacctcccat gtgttcagtt
     2761 cacccatgca ggcatgctga ggtgatgaag aaaatcattg agactgttgc agaaggaggg
     2821 ggagaacttg gagttcatat gtatcttctt attttcttga aatttgtaca agctgtcatt
     2881 ccaacaatag aatatgacta cacaagacac ttcacaatgt aagcggccgc ttgctgagtt
     2941 ggctgctgcc accgctgagc aataactagc ataacccctt ggccgcttcg agcagacatg
     3001 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3061 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3121 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     3181 tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac
     3241 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3301 atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     3361 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3661 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3721 gcaaaaagct tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat
     3781 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3841 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3901 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3961 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4021 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4081 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4141 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4201 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4261 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4321 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4381 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4441 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4501 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4561 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4621 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     4681 tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca
     4741 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     4801 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4861 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4921 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4981 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5041 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5101 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     5161 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5221 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5281 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5341 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5401 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5461 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5521 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5581 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5641 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5701 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5761 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     5821 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6361 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6541 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6661 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     6721 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     6781 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     6841 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     6901 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag
     6961 tactccggcc acgatgcgtc cggcgtagag gatcgagatc ttcaatattg gccattagcc
     7021 atattattca ttggttatat agca
//
