LOCUS       sequence-482812-        7882 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Encodes doubly tagged ACE2 protein. HA on the N-terminal added after
            the signal peptide (a.a 1-17) and FLAG tag at the end of the insert.
            A linker sequence was added after the HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7882)
  TITLE     Zaid Abassi Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ACE2-HA-Linker-FLAG
FEATURES             Location/Qualifiers
     source          1..7882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2395..2418
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2457..2474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2463..2687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2733..3161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2820..2839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3030..3051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3170..3190)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3175..3504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3355..3490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3417..3436
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3625..3644)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4233..4252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4537..4658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4628..4647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4707..4723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4707..4723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4720..4742)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4731..4747
                     /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(4755..4761,4762..4779,4780..4785))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4800..4821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4938..4955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5109..5697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5189..5208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5868..6659,6660..6728))
                     /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     6491..6510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6729..6833)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6908..6927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7099..7478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7479..7682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7633..7653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7727..7746
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7727..7745
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7838..7864
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 cagtccacca ttgaggaaca ggccaagaca tttttggaca agtttaacca cgaagccgaa
       61 gacctgttct atcaaagttc acttgcttct tggaattata acaccaatat tactgaagag
      121 aatgtccaaa acatgaataa tgctggggac aaatggtctg cctttttaaa ggaacagtcc
      181 acacttgccc aaatgtatcc actacaagaa attcagaatc tcacagtcaa gcttcagctg
      241 caggctcttc agcaaaatgg gtcttcagtg ctctcagaag acaagagcaa acggttgaac
      301 acaattctaa atacaatgag caccatctac agtactggaa aagtttgtaa cccagataat
      361 ccacaagaat gcttattact tgaaccaggt ttgaatgaaa taatggcaaa cagtttagac
      421 tacaatgaga ggctctgggc ttgggaaagc tggagatctg aggtcggcaa gcagctgagg
      481 ccattatatg aagagtatgt ggtcttgaaa aatgagatgg caagagcaaa tcattatgag
      541 gactatgggg attattggag aggagactat gaagtaaatg gggtagatgg ctatgactac
      601 agccgcggcc agttgattga agatgtggaa catacctttg aagagattaa accattatat
      661 gaacatcttc atgcctatgt gagggcaaag ttgatgaatg cctatccttc ctatatcagt
      721 ccaattggat gcctccctgc tcatttgctt ggtgatatgt ggggtagatt ttggacaaat
      781 ctgtactctt tgacagttcc ctttggacag aaaccaaaca tagatgttac tgatgcaatg
      841 gtggaccagg cctgggatgc acagagaata ttcaaggagg ccgagaagtt ctttgtatct
      901 gttggtcttc ctaatatgac tcaaggattc tgggaaaatt ccatgctaac ggacccagga
      961 aatgttcaga aagcagtctg ccatcccaca gcttgggacc tggggaaggg cgacttcagg
     1021 atccttatgt gcacaaaggt gacaatggac gacttcctga cagctcatca tgagatgggg
     1081 catatccagt atgatatggc atatgctgca caaccttttc tgctaagaaa tggagctaat
     1141 gaaggattcc atgaagctgt tggggaaatc atgtcacttt ctgcagccac acctaagcat
     1201 ttaaaatcca ttggtcttct gtcacccgat tttcaagaag acaatgaaac agaaataaac
     1261 ttcctgctca aacaagcact cacgattgtt gggactctgc catttactta catgttagag
     1321 aagtggaggt ggatggtctt taaaggggaa attcccaaag accagtggat gaaaaagtgg
     1381 tgggagatga agcgagagat agttggggtg gtggaacctg tgccccatga tgaaacatac
     1441 tgtgaccccg catctctgtt ccatgtttct aatgattact cattcattcg atattacaca
     1501 aggacccttt accaattcca gtttcaagaa gcactttgtc aagcagctaa acatgaaggc
     1561 cctctgcaca aatgtgacat ctcaaactct acagaagctg gacagaaact gttcaatatg
     1621 ctgaggcttg gaaaatcaga accctggacc ctagcattgg aaaatgttgt aggagcaaag
     1681 aacatgaatg taaggccact gctcaactac tttgagccct tatttacctg gctgaaagac
     1741 cagaacaaga attcttttgt gggatggagt accgactgga gtccatatgc agaccaaagc
     1801 atcaaagtga ggataagcct aaaatcagct cttggagata aagcatatga atggaacgac
     1861 aatgaaatgt acctgttccg atcatctgtt gcatatgcta tgaggcagta ctttttaaaa
     1921 gtaaaaaatc agatgattct ttttggggag gaggatgtgc gagtggctaa tttgaaacca
     1981 agaatctcct ttaatttctt tgtcactgca cctaaaaatg tgtctgatat cattcctaga
     2041 actgaagttg aaaaggccat caggatgtcc cggagccgta tcaatgatgc tttccgtctg
     2101 aatgacaaca gcctagagtt tctggggata cagccaacac ttggacctcc taaccagccc
     2161 cctgtttcca tatggctgat tgtttttgga gttgtgatgg gagtgatagt ggttggcatt
     2221 gtcatcctga tcttcactgg gatcagagat cggaagaaga aaaataaagc aagaagtgga
     2281 gaaaatcctt atgcctccat cgatattagc aaaggagaaa ataatccagg attccaaaac
     2341 actgatgatg ttcagacctc ctttggtacc gagacctccc aggtggcgcc cgctgactac
     2401 aaagacgatg acgacaagta agcttggatc cgcgttaagt ttaaaccgct gatcagcctc
     2461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2641 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2701 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2761 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2821 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2881 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2941 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3001 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3061 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3121 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3181 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3361 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3421 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3481 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3541 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3601 tctccggccg cttgggtgga gaggctattc ggctatgact gggcaacaga caatcggctg
     3661 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3721 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3781 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3841 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3901 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3961 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4021 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4081 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4141 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4201 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4261 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4321 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4501 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4621 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4681 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4741 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4801 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4861 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4921 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5161 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5221 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5281 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5341 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5401 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5461 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5521 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5581 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5641 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5701 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5761 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5821 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5881 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5941 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6001 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6061 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6121 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6181 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6241 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6301 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6361 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6421 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6481 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6541 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6601 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6661 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6721 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6781 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6841 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6901 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6961 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7021 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7081 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     7141 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     7201 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     7261 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     7321 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     7381 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     7441 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     7501 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     7561 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     7621 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     7681 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     7741 atagggagac ccaagctggc tagcgtttaa acgggccctc tagaccatgt caagctcttc
     7801 ctggctcctt ctcagccttg ttgctgtaac tgctgcttac ccatacgatg ttccagatta
     7861 cgctggcagc cctggcggtg cc
//
