LOCUS       sequence-145921-        6729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Defective Helper to produce neurotropic, propagation-incompetent 
            Sindbis virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6729)
  TITLE     DH-BB(5'SIN;TE12ORF)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6729)
  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: 
            "Unpublished"
COMMENT     Sequence Label: DH-BB(5'SIN;TE12ORF)
FEATURES             Location/Qualifiers
     source          1..6729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4900..5004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5005..5073,5074..5865)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5223..5242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6036..6624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6525..6544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attgacggcg tagtacacac tattgaatca aacagccgac caattgcact accatcacaa
       61 tggagaagcc agtagtaaac gtagacgtag acccccagag tccgtttgtc gtgcaactgc
      121 aaaaaagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat gaccatgcta
      181 atgccagagc attttcgcat ctggccagta aactaatcga gctggaggtt cctaccacag
      241 cgacgatctt ggacataggc agcgcaccgg ctcgtagaat gttttccgag caccagtatc
      301 attgtgtctg ccccatgcgt agtccagaag acccggaccg catgatgaaa tatgccagta
      361 aactggcgga aaaagcgtgc aagattacaa acaagaactt gcatgagaag attaaggatc
      421 tccgggatcc cctgaaaagg ctgtttaagt tgggtaaacc gctcccagcc gacgacgagc
      481 aagacgaaga cagaagacgc gctctgctag atgaaacaaa ggcgtggttt agagtaggta
      541 taacaggcac tttagcagtg gccgtgacga cccggtatga ggtagacaat attacacctg
      601 tcctactggc attgagaact tttgcccaga gcaaaagagc attccaagcc atcagagggg
      661 aaataaagca tctctacggt ggtcctaaat agtcagcata gtacatttca tctgactaat
      721 actacaacac caccaccatg aatagaggat tctttaacat gctcggccgc cgccccttcc
      781 cggcccccac tgccatgtgg aggccgcgga gaaggaggca ggcggccccg atgcctgccc
      841 gcaacgggct ggcttctcaa atccagcaac tgaccacagc cgtcagtgcc ctagtcattg
      901 gacaggcaac tagacctcaa cccccacgtc cacgcccgcc accgcgccag aagaagcagg
      961 cgcccaagca accaccgaag ccgaagaaac caaaaacgca ggagaagaag aagaagcaac
     1021 ctgcaaaacc caaacccgga aagagacagc gcatggcact taagttggag gccgacagat
     1081 tgttcgacgt caagaacgaa gacggagatg tcatcgggca cgcactggcc atggaaggaa
     1141 aggtaatgaa acctctgcac gtgaaaggaa ccatcgacca ccctgtgcta tcaaagctca
     1201 aatttaccaa gtcgtcagca tacgacatgg agttcgcaca gttgccagtc aacatgagaa
     1261 gtgaggcatt cacctacacc agtgaacacc ccgaaggatt ctataactgg caccacggag
     1321 cggtgcagta tagtggaggt agatttacca tccctcgcgg agtaggaggc agaggagaca
     1381 gcggtcgtcc gatcatggat aactccggtc gggttgtcgc gatagtcctc ggtggcgctg
     1441 atgaaggaac acgaactgcc ctttcggtcg tcacctggaa tagtaaaggg aagacaatta
     1501 agacgacccc ggaagggaca gaagagtggt ccgcagcacc actggtcacg gcaatgtgtt
     1561 tgctcggaaa tgtgagcttc ccatgcgacc gcccgcccac atgctatacc cgcgaacctt
     1621 ccagagccct cgacatcctt gaagagaacg tgaaccatga ggcctacgat accctgctca
     1681 atgccatatt gcggtgcgga tcgtctggca gaagcaaaag aagcgtcact gacgacttta
     1741 ccctgaccag cccctacttg ggcacatgct cgtactgcca ccatactgaa ccgtgcttca
     1801 gccctgttaa gatcgagcag gtctgggacg aagcggacga taacaccata cgcatacaga
     1861 cttccgccca gtttggatac gaccatagcg gagcagcaag cgcaaacaag taccgctaca
     1921 tgtcgcttaa gcaggatcac accgttaaag aaggcaccat ggatgacatc aagattagca
     1981 cctcaggacc gtgtagaagg cttagctaca aaggatactt tctcctcgca aaatgccctc
     2041 caggggacag cgtaacggtt agcatagtga gtagcaactc agcaacgtca tgtacactgg
     2101 cccgcaagat aaaaccaaaa ttcgtgggac gggaaaaata tgatctacct cccgttcacg
     2161 gtaaaaaaat tccttgcaca gtgtacgacc gtctgaaaga aacaactgca ggctacatca
     2221 ctatgcacag gccgggaccg cacgcttata catcctacct ggaagaatca tcagggaaag
     2281 tttacgcaaa gccgccatct gggaagaaca ttacgtatga gtgcaagtgc ggcgactaca
     2341 agaccggaac cgtttcgacc cgcaccgaaa tcactggttg caccgccatc aagcagtgcg
     2401 tcgcctataa gagcgaccaa acgaagtggg tcttcaactc accggacttg atcagacatg
     2461 acgaccacac ggcccaaggg aaattgcatt tgcctttcaa gttgatcccg agtacctgca
     2521 tggtccctgt tgcccacgcg ccgaatgtaa tacatggctt taaacacatc agcctccaat
     2581 tagatacaga ccacttgaca ttgctcacca ccaggagact aggggcaaac ccggaaccaa
     2641 ccactgaatg gatcgtcgga aagacggtca gaaacttcac cgtcgaccga gatggcctgg
     2701 aatacatatg gggaaatcat gagccagtga gggtctatgc ccaagagtca gcaccaggag
     2761 accctcacgg atggccacac gaaatagtac agcattacta ccatcgccat cctgtgtaca
     2821 ccatcttagc cgtcgcatca gctaccgtgg cgatgatgat tggcgtaact gttgcagtgt
     2881 tatgtgcctg taaagcgcgc cgtgagtgcc tgacgccata cgccctggcc ccaaacgccg
     2941 taatcccaac ttcgctggca ctcttgtgct gcgttaggtc ggccaatgct gaaacgttca
     3001 ccgagaccat gagttacttg tggtcgaaca gtcagccgtt cttctgggtc cagttgtgca
     3061 tacctttggc cgctttcatc gttctaatgc gctgctgctc ctgctgcctg ccttttttag
     3121 tggttgccgg cgcctacctg gcgaaggtag acgcctacga acatgcgacc actgttccaa
     3181 atgtgccaca gataccgtat aaggcacttg ttgaaagggc agggtatgcc ccgctcaatt
     3241 tggagatcac tgtcatgtcc tcggaggttt tgccttccac caaccaagag tacattacct
     3301 gcaaattcac cactgtggtc ccctccccaa aaatcaaatg ctgcggctcc ttggaatgtc
     3361 agccggccgc tcatgcagac tatacctgca aggtcttcgg aggggtctac ccctttatgt
     3421 ggggaggagc gcaatgtttt tgcgacagtg agaacagcca gatgagtgag gcgtacgtcg
     3481 aattgtcagc agattgcgcg tctgaccacg cgcaggcgat taaggtgcac actgccgcga
     3541 tgaaagtagg actgcgtata gtgtacggga acactaccag tttcctagat gtgtacgtga
     3601 acggagtcac accaggaacg tctaaagact tgaaagtcat agctggacca atttcagcat
     3661 catttacgcc attcgatcat aaggtcgtta tccatcgcgg cctggtgtac aactatgact
     3721 tcccggaata tggagcgatg aaaccaggag cgtttggaga cattcaagct acctccttga
     3781 ctagcaagga tctcatcgcc agcacagaca ttaggctact caagccttcc gccaagaatg
     3841 tgcatgtccc gtacacgcag gccgcatcag gatttgagat gtggaaaaac aactcaggcc
     3901 gcccattgca ggaaaccgca cctttcgggt gtaagattgc agtaaatccg ctccgagcgg
     3961 tggactgttc atacgggaac attcccattt ctattgacat cccgaacgct gcctttatca
     4021 ggacatcaga tgcaccactg gtctcaacag tcaaatgtga agtcagtgag tgcacttatt
     4081 cagcagactt cgacgggatg gccaccctgc agtatgtatc cgaccgcgaa ggtcaatgcc
     4141 ccgtacattc gcattcgagc acagcaactc tccaagagtc gacagtacat gtcctggaga
     4201 aaggagcggt gacagtacac tttagcaccg cgagtccaca ggcgaacttt atcgtatcgc
     4261 tgtgtgggaa gaagacaaca tgcaatgcag aatgtaaacc accagctgac catatcgtga
     4321 gcaccccgca caaaaatgac caagaatttc aagccgccat ctcaaaaaca tcatggagtt
     4381 ggctgtttgc ccttttcggc ggcgcctcgt cgctattaat tataggactt atgatttttg
     4441 cttgcagcat gatgctgact agcacacgaa gatgaccgct acgccccaat gatccgacca
     4501 gcaaaactcg atgtacttcc gaggaactga tgtgcataat gcatcaggct ggtacattag
     4561 atccccgctt accgcgggca atatagcaac actaaaaact cgatgtactt ccgaggaagc
     4621 gcagtgcata atgctgcgca gtgttgccac ataaccacta tattaaccat ttatctagcg
     4681 gacgccaaaa actcaatgta tttctgagga agcgtggtgc ataatgccac gcagcgtctg
     4741 cataactttt attatttctt ttattaatca acaaaatttt gtttttaaca tttcaaaaaa
     4801 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agggaattcc tcgaggggaa ttaattcttg
     4861 aagacgaaag ggccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4921 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4981 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5041 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5101 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5161 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5221 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5281 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5341 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5401 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5461 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5521 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5581 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5641 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5701 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5761 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5821 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5881 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6181 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6421 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcga gctcgtatgg acatattgtc
     6661 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttagggg
     6721 acactatag
//
