LOCUS       sequence-261097-        6817 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 6817)
  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 6817)
  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..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..222
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2600..2641
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2723..2764
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2774..2791
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2814..2831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2820..3044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3090..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3177..3196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3387..3408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3527..3547)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3532..3861
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3712..3847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3774..3793
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3909..3956
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3975..4349
                     /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    4479..4600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4516..4535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4570..4589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4649..4665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4649..4665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4662..4684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4673..4689
                     /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(4697..4703,4704..4721,4722..4727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4742..4763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5810..6601,6602..6670))
                     /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     6433..6452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6671..6775)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agatcgtacc gagctcttac gcgggtcgaa gcggagtact gtcctccgag tggagtactg
       61 tcctccgagc ggagtactgt cctccgagtc gagggtcgaa gcggagtact gtcctccgag
      121 tggagtactg tcctccgagc ggagtactgt cctccgagtc gactctagag ggtatataat
      181 ggatctcgag atatcggagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
      241 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg
      301 tgcattggaa cgcgcattcc ccgtgttaat taacaggtaa gtgtcttcct cctgtttcct
      361 tcccctgcta ttctgctcaa ccttcctatc agaaactgca gtatctgtat ttttgctagc
      421 agtaatacta acggttcttt ttttctcttc acaggccacc aagcttggta ccatggtgaa
      481 cgcgatttta atggcttgta tactcgccgg aattttcgtc ggaatgttca acgaagtcga
      541 cggaattcag attttatcga aatcaaagct cgaaaagtgt gagaaaacat cagattctgg
      601 taatctcaat tgttccacta agatcgtctt gaatctcgct gttcccagtg gatccagtgg
      661 aggagaggct tcgattgtag cggagatcgt ggaggttgaa gataactcga gtagtaatat
      721 gcaaaccgta cggattccac cggttattac agtgaacaaa tctgcagcat atgctctcta
      781 tgatcttact tacattaggg atgtacctta taaacctcaa gagtatcatg tcacgactcg
      841 aaagtgtgaa cccgatgctg gaccggatat tgtacaaatt tgtgagaggt tgagagatga
      901 gaagggtaat gttttagagc agactcagcc gatttgttgt ccatgtggac cgcaacgaag
      961 aatgccttct tcttgtggag atatttttga taagatgatt aaagggaagg caaataccgc
     1021 gcattgcctt cgttttccag gtgactggtt ccatgttttc gggattggac aacggtcact
     1081 aggatttagt gtgcgagttg agttgaaaac aggaactaga gtttcagaag tgatcattgg
     1141 tcctgaaaat aggacagcta ctgcaaatga taactttctc aaagttaatc tcataggaga
     1201 ctttggtggt tacacaagta tcccttcttt cgaggacttc tatcttgtaa tcccaagaga
     1261 ggcggctgaa gcgggacaac cagggagctt aggcgcaaat tactcaatgt ggatgctgct
     1321 tgagagagtg agatttacat tggatggctt agaatgtaat aaaatcggtg ttggttatga
     1381 agctttcaac acccaaccaa atttctgctc ttctccatat tggagttgct tgcataatca
     1441 actatggaac ttccgtgagt cagatataaa cagaatcgac cgacaccaac ttcctttata
     1501 tggacttgaa ggaagatttg agaggatcaa tcaacaccca aacgcggggc ctcactcgtt
     1561 ctcaattgga gtcacggaaa cactcaacac aaatctaatg atagaactaa gagctgatga
     1621 tattgaatac gttttccaga ggagtcccgg taagatcata aacatcgcga taccgacatt
     1681 cgaagcactt acacaattcg gagttgctgc agtcataatc aagaatactg gtgaagtaga
     1741 agcgtcttat agcttaacgt ttgattgctc caaaggcgtg gcttttgttg aggaacaatt
     1801 tttcattata aaaccgaaag cggttactac tcggtctttt aagctgtacc cgacgaaaga
     1861 tcaggctgca aagtatatat gcacagctat actcaaggac tctcagttca gcgaagtaga
     1921 ccgagccgag tgtcaattta gtacaacggc taccgtactt gacaatggaa ctcaggttac
     1981 taatccattt caaattccag agactcaacc taaaggtttc tttgattcga tcaggatact
     2041 atggatgaaa atcatcaatg gtttagtcga tttcatcacc ggagatactt gcagaaacaa
     2101 atgctcgagt ttcttcgact tcagttgcca cattcaatac gtttgcttaa gctggatggt
     2161 tatgtttggt cttctcctcg ctctcttccc aattacttgt ttgcttcttt ggctcttgca
     2221 ccaaaaggga ctctttgatc catgctacga ctggtgggaa gatcatttcg atttagacca
     2281 tcatcggaga ttattaccct ccagagcaga cgttgttaac cgccatcatc accatcacaa
     2341 gcatcgtcat caccataatc accatcgtag gacacaccag agacacaaac atcatcatgg
     2401 tcaagacgac gacgtattgc agaagatgat gcttgaacgt gaccacagcg actctcacta
     2461 ctaccaccaa cttcatcgag tccataaaga tagcaagcag aagcagcgta ggcgagccaa
     2521 gcacgggatc gtcctgccac gtgatgtgca cgtggaaaga cagaggaaac aaagactacg
     2581 tgagagtccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2641 gtgaactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag
     2701 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     2761 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     2821 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2881 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2941 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3001 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3061 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3241 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3421 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3541 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3661 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3781 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3841 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3901 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3961 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     4021 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     4081 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     4141 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     4201 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     4261 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     4321 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     4381 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4441 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4621 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6781 ttccccgaaa agtgccacct gacgtcgacg gatcggg
//
