LOCUS       sequence-256644-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express AtHAP2 full length protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6723)
  AUTHORS   Valansi C, Moi D, Leikina E, Matveev E, Grana M, Chernomordik LV, 
            Romero H, Aguilar PS, Podbilewicz B
  TITLE     Arabidopsis HAP2/GCS1 is a gamete fusion protein homologous to 
            somatic and viral fusogens.
  JOURNAL   J Cell Biol. 2017 Mar 6;216(3):571-581. doi: 10.1083/jcb.201610093. 
            Epub 2017 Jan 30.
  PUBMED    28137780
REFERENCE   2  (bases 1 to 6723)
  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: 
            "10.1083/jcb.201610093"; journalName: "J Cell Biol"; date: 
            "2017-03-6- 6"; volume: "216"; issue: "3"; pages: "571-581"
COMMENT     Sequence Label: pGENE B::AtHAP2-V5
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..214
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2624..2665
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2675..2692
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2715..2732)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2721..2945
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2991..3419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3078..3097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3288..3309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3428..3448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3433..3762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3613..3748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3675..3694
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3810..3857
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3876..4250
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4380..4501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4417..4436)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4471..4490
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4550..4566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4550..4566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4563..4585)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4574..4590
                     /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)."
     promoter        complement(join(4598..4604,4605..4622,4623..4628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4643..4664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4781..4798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4952..5537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5032..5051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5708..6499,6500..6568))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYATGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6331..6350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6569..6673)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgagctctt acgcgggtcg aagcggagta ctgtcctccg agtggagtac tgtcctccga
       61 gcggagtact gtcctccgag tcgagggtcg aagcggagta ctgtcctccg agtggagtac
      121 tgtcctccga gcggagtact gtcctccgag tcgactctag agggtatata atggatctcg
      181 agatatcgga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
      241 tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac ggtgcattgg
      301 aacgcgcatt ccccgtgtta attaacaggt aagtgtcttc ctcctgtttc cttcccctgc
      361 tattctgctc aaccttccta tcagaaactg cagtatctgt atttttgcta gcatggtgaa
      421 cgcgatttta atggcttgta ttcttgctgg aattttcgtt ggaatgttca atgaagttga
      481 tggaattcaa attctttcta aatctaaact tgaaaaatgt gaaaaaactt ctgattctgg
      541 aaatcttaat tgttctacta aaattgttct taatcttgct gttccatctg gatcttctgg
      601 aggagaagct tctattgttg ctgaaattgt tgaagttgaa gataattctt cttctaatat
      661 gcaaactgtt agaattccac cagttattac tgttaataaa tctgctgctt atgctcttta
      721 tgatcttact tatattagag atgttccata taaaccacaa gaatatcatg ttactactag
      781 aaaatgtgaa ccagatgctg gaccagatat tgttcaaatt tgtgaaagac ttagagatga
      841 aaaaggaaat gttcttgaac aaactcaacc aatttgttgt ccatgtggac cacaaagaag
      901 aatgccatct tcttgtggag atattttcga taaaatgatt aaaggaaaag ctaatactgc
      961 tcattgtctt agattcccag gagattggtt ccatgttttc ggaattggac aaagatctct
     1021 tggattctct gttagagttg aacttaaaac tggaactaga gtttctgaag ttattattgg
     1081 accagaaaat agaactgcta ctgctaatga taatttcctt aaagttaatc ttattggaga
     1141 tttcggagga tatacttcta ttccatcttt cgaagatttc tatcttgtta ttccaagaga
     1201 agctgctgaa gctggacaac caggatctct tggagctaat tattctatgt ggatgcttct
     1261 tgaaagagtt agattcactc ttgatggact tgaatgtaat aaaattggag ttggatatga
     1321 agctttcaat actcaaccaa atttctgttc ttctccatat tggtcttgtc ttcataatca
     1381 actttggaat ttcagagaat ctgatattaa tagaattgat agacatcaac ttccacttta
     1441 tggacttgaa ggaagattcg aaagaattaa tcaacatcca aatgctggac cacattcttt
     1501 ctctattgga gttactgaaa ctcttaatac taatcttatg attgaactta gagctgatga
     1561 tattgaatat gttttccaaa gatctccagg aaaaattatt aatattgcta ttccaacttt
     1621 cgaagctctt actcaattcg gagttgctgc tgttattatt aaaaatactg gagaagttga
     1681 agcttcttat tctcttactt tcgattgttc taaaggagtt gctttcgttg aagaacaatt
     1741 cttcattatt aaaccaaaag ctgttactac tagatctttc aaactttatc caactaaaga
     1801 tcaagctgct aaatatattt gtactgctat tcttaaagat tctcaattct ctgaagttga
     1861 tagagctgaa tgtcaattct ctactactgc tactgttctt gataatggaa ctcaagttac
     1921 taatccattc caaattccag aaactcaacc aaaaggattc ttcgattcta ttagaattct
     1981 ttggactaaa attattaatg gacttgttga tttcattact ggagatactt gtagaaataa
     2041 atgttcttct ttcttcgatt tctcttgtca tattcaatat gtttgtcttt cttggatggt
     2101 tatgttcgga cttcttcttg ctcttttccc aattacttgt cttcttcttt ggcttcttca
     2161 tcaaaaagga cttttcgatc catgttatga ttggtgggaa gatcatttcg atcttgatca
     2221 tcatagaaga cttcttccat ctagagctga tgttgttaat agacatcatc atcatcataa
     2281 acatagacat catcataatc atcatagaag aactcatcaa agacataaac atcatcatgg
     2341 acaagatgat gatgttcttc aaaaaatgat gcttgaaaga gatcattctg attctcatta
     2401 ttatcatcaa cttcatagag ttcataaaga ttctaaacaa aaacaaagaa gaagagctaa
     2461 acatggaatt gttcttccaa gagatgttca tgttgaaaga caaaggaaac aaagactacg
     2521 tgagagtggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
     2581 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     2641 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
     2701 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2761 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2821 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2881 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2941 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3001 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3061 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3121 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3181 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3241 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3301 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3361 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3421 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3661 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3781 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3841 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     3901 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     3961 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4021 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4081 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4141 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4201 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     4261 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4321 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4441 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4501 atcatgtctg tataccgtcg acatctagct agagcttggc gtaatcatgg tcatagctgt
     4561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5461 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5761 catccatagt tgcctgactc cccgtcgcgt agataactac gatacgggag ggcttaccat
     5821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6721 gta
//
