LOCUS       sequence-213248-        5232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of HA ZNHIT3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5232)
  AUTHORS   Wyant GA, Abu-Remaileh M, Frenkel EM, Laqtom NN, Dharamdasani V, 
            Lewis CA, Chan SH, Heinze I, Ori A, Sabatini DM
  TITLE     NUFIP1 is a ribosome receptor for starvation-induced ribophagy.
  JOURNAL   Science. 2018 May 18;360(6390):751-758. doi: 
            10.1126/science.aar2663. Epub 2018 Apr 26.
  PUBMED    29700228
REFERENCE   2  (bases 1 to 5232)
  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"; doi: 
            "10.1126/science.aar2663"; journalName: "Science"; date: 
            "2018-05-18- 18"; volume: "360"; issue: "6390"; pages: "751-758"
COMMENT     Sequence Label: pRK5-HA ZNHIT3
FEATURES             Location/Qualifiers
     source          1..5232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        435..453
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     435..452
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     466..485
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      570..579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             582..608
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1139..1148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1152..1286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1189..1208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1243..1262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1355..1712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1355..1546
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1378..1398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1563..1698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1732..1749)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1732..1748)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1741..1763)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1961..2416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2048..2067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2258..2279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2433..2452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2552..2574
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2612..2630)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2698..2802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2803..2871,2872..3663)
                     /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(3021..3040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3834..4422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4323..4342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4576..4593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4710..4731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4746..4751,4752..4769,4770..4776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4784..4800
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4789..4811
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4808..4824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4808..4824
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        4853..5232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      301 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      361 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
      421 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
      481 caggtgtcca ctcccaggtc caactgcacc tcggttctat cgattgaatt cccgaaccga
      541 cagtcggtct cttcaccaag gccattcgcg ccaccatggg ctacccctat gatgtgcctg
      601 actacgcaga tctcaatggt ggtggtggtg ggtcgacgat ggcgtcgctc aaatgtagca
      661 ccgtcgtctg cgtgatctgc ttggagaagc ccaaataccg ctgtccagcc tgccgcgtgc
      721 cctactgctc ggtagtctgc ttccggaagc acaaagaaca gtgcaaccct gaaactcgtc
      781 ctgttgagaa aaaaataaga tcagctcttc ctaccaaaac cgtaaagcct gtggaaaaca
      841 aagatgatga tgactctata gctgattttc tcaatagtga tgaggaagaa gacagagttt
      901 ctttgcagaa tttaaagaat ttaggggaat ctgcaacatt aagaagctta ttgctcaatc
      961 cacacctcag gcagttgatg gtcaacctcg atcagggaga agacaaagca aagctcatga
     1021 gagcttacat gcaagagcct ttgtttgtgg agtttgcaga ctgctgttta ggaattgtgg
     1081 agccatccca gaatgaggag tcttagcggc cgctaagtaa gtaaggatcc ccagcttggc
     1141 cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1201 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1261 tcaatgtatc ttatcatgtc tggatcggga attaattcgg cgcagcacca tggcctgaaa
     1321 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     1381 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1441 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     1501 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1561 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1621 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1681 gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg cactggccgt
     1741 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1801 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1861 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     1921 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     1981 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2041 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2101 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2161 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2221 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2281 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2341 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2401 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2461 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2521 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2581 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2641 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2701 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2761 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2821 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2881 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2941 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3001 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3061 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3481 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3661 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3721 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3961 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4141 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4441 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4561 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4621 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4681 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4741 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4801 tttcacacag gaaacagcta tgacatgatt acgaattaat tcgagctcgc ccgacattga
     4861 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4921 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4981 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5041 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5101 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5161 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     5221 gctattacca tg
//
