LOCUS       sequence-407377-        7341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of env polyprotein from equine infectious anaemia (EIAV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 7341)
  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"; journalName: "Nat 
            Commun."; date: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: pHCMV_EIAV_Env
FEATURES             Location/Qualifiers
     source          1..7341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1419..1428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     RBS             1622..1633
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4046..4065)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4111..4166
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4165..4184)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4524..4540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4524..4540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4537..4559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4548..4564
                     /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(4572..4578,4579..4596,4597..4602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4617..4638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4755..4772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4926..5514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5006..5025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5685..6476,6477..6545))
                     /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     6308..6327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6546..6650)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6676..7131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6813..6834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7025..7044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7258..7280
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7272..7289
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7273..7289
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7299..7318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7299..7317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaca cgccatccac
      721 gctgttttga cctccataga tgacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcgc caccatggtc tccatcgcct
     1441 tctacggcgg catccccgga ggcatctcca cacctatcac acagcagagc gagaagagca
     1501 aatgtgagga gaacaccatg ttccaaccct attgctacaa taacgattcc aagaactcca
     1561 tggctgagag caaggaagct agagatcaag agatgaatct gaaagaggaa agcaaggagg
     1621 aaaagaggag aaatgactgg tggaagatcg gcatgtttct gctgtgtctg gccggaacca
     1681 ccggcggcat tctgtggtgg tacgagggcc tcccccaaca gcactatatc ggactggtgg
     1741 ccattggcgg aaggctcaat ggcagcggcc agtccaacgc catcgaatgt tggggcagct
     1801 tccccggctg tagacccttt cagaactatt tcagctacga gaccaataga agcatgcaca
     1861 tggacaataa caccgctaca ctgctcgagg cttaccatag ggaaatcacc ttcatttaca
     1921 agagcagctg caccgacagc gaccactgtc aagagtacca gtgcaaaaag gtgaatctga
     1981 acagctccga tagcagcaac agcgtcagag tcgaggatgt gatgaacacc gccgaatact
     2041 ggggcttcaa atggctcgag tgcaaccaga cagagaactt caagaccatt ctggtccccg
     2101 agaatgaaat ggtgaacatt aacgataccg acacatggat ccctaagggc tgtaacgaga
     2161 catgggctag agtcaagaga tgccctattg acattctgta cggcatccat cccatcagac
     2221 tgtgcgtgca gccccccttt tttctggtgc aagagaaggg catcgctgac accagcagaa
     2281 tcggaaattg cggccccacc atttttctgg gagtgctcga ggataacaag ggcgtggtga
     2341 ggggcgatta caccgcttgc aacgtgagga gactgaacat taatagaaag gattataccg
     2401 gcatttacca agtgcccatt ttctacacat gcaccttcac caatattacc agctgcaata
     2461 atgagcctat catcagcgtc atcatgtatg agaccaacca agtgcagtat ctgctgtgca
     2521 ataacaacaa ctccaataac tataactgtg tcgtgcagtc cttcggcgtc attggccaag
     2581 cccatctgga actccctaga cctaataaga ggattagaaa ccagtccttt aatcagtaca
     2641 attgcagcat caataataag accgagctcg agacatggaa actcgtcaag accagcggca
     2701 tcacccctct gcccatttcc agcgaagcca acaccggact gattagacac aagagggact
     2761 tcggcatttc cgccattgtg gccgccattg tcgccgctac cgctatcgct gcttccgcta
     2821 caatgtccta cgtggccctc accgaggtga acaaaatcat ggaagtccag aaccacacct
     2881 tcgaggtgga aaactccaca ctgaacggca tggacctcat tgaaagacag attaagattc
     2941 tgtacgccat gattctgcag acccacgctg acgtgcagct cctcaaggaa aggcaacaag
     3001 tggaggagac ctttaacctc atcggctgta tcgagaggac acatgtgttc tgccataccg
     3061 gccatccttg gaatatgagc tggggccatc tgaacgagag cacccagtgg gacgattggg
     3121 tctccaaaat ggaggacctc aaccaagaga ttctgacaac actgcatgga gccagaaaca
     3181 acctcgccca gtccatgatc accttcaaca cccccgactc catcgcccag tttggcaagg
     3241 atctgtggag ccatatcggc aactggatcc ccggcctcgg cgccagcatc attaagtata
     3301 ttgtcatgtt tctgctcatc tatctgctgc tgaccagctc ccctaagatt ctgagagccc
     3361 tctggaaagt gacatccggc gctggaagca gcggatctag atatctcaag aagaagtttc
     3421 accataagca cgctagcaga gaggacacat gggaccaagc ccagcataac atccatctcg
     3481 ctggcgtcac cggcggaagc ggcgacaaat attacaaaca gaagtattct agaaatgatt
     3541 ggaacggcga gagcgaggag tacaacagaa ggcccaagag ctgggtgaag agcattgagg
     3601 ccttcggcga gtcctacatc agcgagaaaa ccaagggaga gattagccaa cccggcgccg
     3661 ctatcaatga gcacaagaac ggctccggcg gaaacaaccc ccaccaaggc tctctggatc
     3721 tcgagatcag aagcgaggga ggcaacatct acgactgttg catcaaagct caagagggca
     3781 cactggctat cccttgctgc ggcttccctc tgtggctctt ttggggcctc gtcattattg
     3841 tcggcagaat cgccggctat ggactgaggg gactcgccgt cattattaga atttgcatta
     3901 gaggactgaa tctgatcttc gagatcatca gaaagatgct ggactatatc ggaagggccc
     3961 tcaatcccgg aacatcccac gtcagcatgc ctcagtatgt ctgatgtctt cctggagctc
     4021 cagtcattga ctataacgtc tcagatcttt ttccctctgc caaaaattat ggggacatca
     4081 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4141 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4201 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     4261 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     4321 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     4381 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4441 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacg
     4501 gatcggccgc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5101 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5341 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5821 ccgcgagacc cacgctcacc ggcaccagat ttatcagcaa taaaccagcc agccggaagg
     5881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6661 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     6721 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     6781 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     6841 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     6901 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     6961 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7021 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     7081 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     7141 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7201 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     7261 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     7321 aattggagct ccaccgcggt g
//
