LOCUS       sequence-59801-e       10249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10249)
  AUTHORS   Patel M, Giddings AM, Sechelski J, Olsen JC
  TITLE     High efficiency gene transfer to airways of mice using influenza 
            hemagglutinin pseudotyped lentiviral vectors.
  JOURNAL   J Gene Med. 2013 Jan;15(1):51-62. doi: 10.1002/jgm.2695.
  PUBMED    23319179
REFERENCE   2  (bases 1 to 10249)
  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: "10.1002/jgm"; 
            journalName: "J Gene Med"; date: "2013-01"; volume: "15"; issue: 
            "1"; pages: "51-62"
COMMENT     Sequence Label: pEV53D
FEATURES             Location/Qualifiers
     source          1..10249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7464..7481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7470..7694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7827..7846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7922..8043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7959..7978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8013..8032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8092..8108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8092..8108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8105..8127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8116..8132
                     /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(8140..8146,8147..8164,8165..8170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8185..8206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8323..8340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8494..9082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8574..8593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9253..10044,10045..10113))
                     /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     9876..9895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10114..10218)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 cgcgggcgcg ccaggtaaga tgggagaccc tttgacatgg agcaaggcgc tcaagaagtt
      961 agagaaggtg acggtacaag ggtctcagaa attaactact ggtaactgta attgggcgct
     1021 aagtctagta gacttatttc atgataccaa ctttgtaaaa gaaaaggact ggcagctgag
     1081 ggatgtcatt ccattgctgg aagatgtaac tcagacgctg tcaggacaag aaagagaggc
     1141 ctttgaaaga acatggtggg caatttctgc tgtaaagatg ggcctccaga ttaataatgt
     1201 agtagatgga aaggcatcat tccagctcct aagagcgaaa tatgaaaaga agactgctaa
     1261 taaaaagcag tctgagccct ctgaagaata tccaatcatg atagatggga ctggaaacag
     1321 aaattttaga cctctaacac ctagaggata tactacttgg gtgaatacca tacagacaaa
     1381 tggtctatta aatgaagcta gtcaaaactt atttgggata ttatcagtag actgtacttc
     1441 tgaagaaatg aatgcatttt tggatgtggt acctggccag gcaggacaaa agcagatatt
     1501 acttgatgca attgataaaa tagcagatga ttgggataat agacatccat taccgaatgc
     1561 tccactggtg gcaccaccac aagggcctat tcccatgaca gcaaggttta ttagaggttt
     1621 aggagtacct agagaaagac agatggagcc tgcttttgat cagtttaggc agacatatag
     1681 acaatggata atagaagcca tgtcagaagg catcaaagtg atgattggaa aacctaaagc
     1741 tcaaaatatt aggcaaggag ctaaggaacc ttacccagaa tttgtagaca gactattatc
     1801 ccaaataaaa agtgagggac atccacaaga gatttcaaaa ttcttgactg atacactgac
     1861 tattcagaac gcaaatgagg aatgtagaaa tgctatgaga catttaagac cagaggatac
     1921 attagaagag aaaatgtatg cttgcagaga cattggaact acaaaacaaa agatgatgtt
     1981 attggcaaaa gcacttcaga ctggtcttgc gggcccattt aaaggtggag ccttaaaagg
     2041 agggccacta aaggcagcac aaacatgtta taactgtggg aagccaggac atttatctag
     2101 tcaatgtaga gcacctaaag tctgttttaa atgtaaacag cctggacatt tctcaaagca
     2161 atgcagaagt gttccaaaaa acgggaagca aggggctcaa gggaggcccc agaaacaaac
     2221 tttcccgata caacagaaga gtcagcacaa caaatctgtt gtacaagaga ctcctcagac
     2281 tcaaaatctg tacccagatc tgagcgaaat aaaaaaggaa tacaatgtca aggagaagga
     2341 tcaagtagag gatctcaacc tggacagttt gtgggagtaa catataatct agagaaaagg
     2401 cctactacaa tagtattaat taatgatact cccttaaatg tactgttaga cacaggagca
     2461 gatacttcag tgttgactac tgcacattat aataggttaa aatatagagg gagaaaatat
     2521 caagggacgg gaataatagg agtgggagga aatgtggaaa cattttctac gcctgtgact
     2581 ataaaggaaa agggtagaca cattaagaca agaatgctag tggcagatat tccagtgact
     2641 attttgggac gagatattct tcaggactta ggtgcaaaat tggttttggc acagctctcc
     2701 aaggaaataa aatttagaaa aatagagtta aaagagggca caatggggcc aaaaattcct
     2761 caatggccac tcactaagga gaaactagaa ggggctaaag agatagtcca aagactattg
     2821 tcagagggaa aaatatcaga agctagtgac aataatcctt ataattcacc catatttgta
     2881 ataaaaaaga ggtctggcaa atggaggtta ttacaagatc tgagagaatt aaacaaaaca
     2941 gtacaagtag gaacggaaat atccagagga ttgcctcacc cgggaggatt aattaaatgt
     3001 aaacacatga ctgtattaga tattggagat gcatatttca ctataccctt agatccagag
     3061 tttagaccat atacagcttt cactattccc tccattaatc atcaagaacc agataaaaga
     3121 tatgtgtgga attgtttacc acaaggattc gtgttgagcc catatatata tcagaaaaca
     3181 ttacaggaaa ttttacaacc ttttagggaa agatatcctg aagtacaatt gtatcaatat
     3241 atggatgatt tgttcgtggg aagtaatggt tctaaaaaac aacacaaaga gttaatcata
     3301 gaattaaggg caatcttact ggaaaagggt tttgagacac cagatgataa attacaagaa
     3361 gtgccacctt atagctggct aggttatcaa ctttgtcctg aaaattggaa agtacaaaaa
     3421 atgcaattag acatggtaaa gaatccaacc cttaatgatg tgcaaaaatt aatggggaat
     3481 ataacatgga tgagctcagg ggtcccaggg ttgacagtaa aacacatagc agctactact
     3541 aagggatgtt tagagttgaa tcaaaaagta atttggacgg aagaggcaca aaaagagtta
     3601 gaagaaaata atgagaagat taaaaatgct caagggttac aatattataa tccagaagaa
     3661 gaaatgttat gtgaggttga aattacaaaa aattatgagg caacttatgt tataaaacaa
     3721 tcacaaggaa tcctatgggc aggtaaaaag attatgaagg ctaataaggg atggtcaaca
     3781 gtaaaaaatt taatgttact gttgcaacat gtggcaacag aaagtattac tagagtagga
     3841 aaatgtccaa cgtttaaggt accatttacc aaagagcaag taatgtggga aatgcaaaaa
     3901 ggatggtatt attcttggct cccagaaata gtatatacac atcaagtagt tcatgatgat
     3961 tggagaatga aattggtaga agaacctaca tcaggaataa caatatacac tgatggggga
     4021 aaacaaaatg gagaaggaat agcagcttat gtgaccagta atgggagaac taaacagaaa
     4081 aggttaggac ctgtcactca tcaagttgct gaaagaatgg caatacaaat ggcattagag
     4141 gataccagag ataaacaagt aaatatagta actgatagtt attattgttg gaaaaatatt
     4201 acagaaggat taggtttaga aggaccacaa agtccttggt ggcctataat acaaaatata
     4261 cgagaaaaag agatagttta ttttgcttgg gtacctggtc acaaagggat atgtggtaat
     4321 caattggcag atgaagccgc aaaaataaaa gaagaaatca tgctagcata ccaaggcaca
     4381 caaattaaag agaaaagaga tgaagatgca gggtttgact tatgtgttcc ttatgacatc
     4441 atgatacctg tatctgacac aaaaatcata cccacagatg taaaaattca agttcctcct
     4501 aatagctttg gatgggtcac tgggaaatca tcaatggcaa aacaggggtt attaattaat
     4561 ggaggaataa ttgatgaagg atatacagga gaaatacaag tgatatgtac taatattgga
     4621 aaaagtaata ttaaattaat agagggacaa aaatttgcac aattaattat actacagcat
     4681 cactcaaatt ccagacagcc ttgggatgaa aataaaatat ctcagagagg ggataaagga
     4741 tttggaagta caggagtatt ctgggtagaa aatattcagg aagcacaaga tgaacatgag
     4801 aattggcata catcaccaaa gatattggca agaaattata agataccatt gactgtagca
     4861 aaacagataa ctcaagaatg tcctcattgc actaagcaag gatcaggacc tgcaggttgt
     4921 gtcatgagat ctcctaatca ttggcaggca gattgcacac atttggacaa taagataata
     4981 ttgacttttg tagagtcaaa ttcaggatac atacatgcta cattattgtc aaaagaaaat
     5041 gcattatgta cttcattggc tattttagaa tgggcaagat tgttttcacc aaagtcctta
     5101 cacacagata acggcactaa ttttgtggca gaaccagttg taaatttgtt gaagttccta
     5161 aagatagcac ataccacagg aataccatat catccagaaa gtcagggtat tgtagaaagg
     5221 gcaaatagga ccttgaaaga gaagattcaa agtcatagag acaacactca aacactggag
     5281 gcagctttac aacttgctct cattacttgt aacaaaggga gggaaagtat gggaggacag
     5341 acaccatggg aagtatttat cactaatcaa gcacaagtaa tacatgagaa acttttacta
     5401 cagcaagcac aatcctccaa aaaattttgt ttttacaaaa tccctggtga acatgattgg
     5461 aagggaccta ctagggtgct gtggaagggt gatggtgcag tagtagttaa tgatgaagga
     5521 aagggaataa ttgctgtacc attaaccagg actaagttac taataaagcc aaattgagta
     5581 ttgttgcagg aagcaagacc caactaccat tgtcagctgt gtttcctgag gtctctagga
     5641 attgattacc tcgatgcttc attaaggaag aagaataaac aaagactgaa ggcaatccaa
     5701 caaggaagac aacctcaata tttgttataa ggtttgatat atgggattat ttggtaaagg
     5761 ggtaacatgg tcagcatcgc attctatggg gggatcccag ggggaatctc aacccctatt
     5821 acccaacagt cagaaaaatc taagtgtgag gagaacacaa tgtttcaacc ttattgttat
     5881 aataatgaca gtaagaacag catggcagaa tcgaaggaag caagagacca agaaatgaac
     5941 ctgaaagaag aatctaaaga agaaaaaaga agaaatgact ggtggaaaaa aggtatgttt
     6001 ctgttatgct tagcaggaac tactggagga atactttggt ggtatgaagg actcccacag
     6061 caacattata tagggttggt ggcgataggg ggaagattaa acggatctgg ccaatcaaat
     6121 gctatagaat gctggggttc cttcccgggg tgtagaccat ttcaaaatta cttcagttat
     6181 gagaccaata gaagcatgca tatggataat aatactgcta cattattaga agctttaacc
     6241 aatataactg ctctataaat aacaaaacag aattagaaac atggaagtta gtaaagactt
     6301 ctggcgtaac tcctttacct atttcttctg aagctaacac tggactaatt agacataaga
     6361 gagattttgg tataagtgca atagtggcag ctattgtagc cgctactgct attgctgcta
     6421 gcgctactat gtcttatgtt gctctaactg aggttaacaa aataatggaa gtacaaaatc
     6481 atacttttga ggtagaaaat agtactctaa atggtatgga tttaatagaa cgacaaataa
     6541 agatattata tgctatgatt cttcaaacac atgcagatgt tcaactgtta aaggaaagac
     6601 aacaggtaga ggagacattt aatttaattg gatgtataga aagaacacat gtattttgtc
     6661 atactggtca tccctggaat atgtcatggg gacatttaaa tgagtcaaca caatgggatg
     6721 actgggtaag caaaatggaa gatttaaatc aagagatact aactacactt catggagcta
     6781 ggaacaattt ggcacaatcc atgataacat tcaatacacc agatagtata gctcaatttg
     6841 gaaaagacct ttggagtcat attggaaatt ggattcctgg attgggagct tccattataa
     6901 aatatatagt gatgtttttg cttatttatt tgttactaac ctcttcgcct aagatcctca
     6961 gggccctctg gaaagtgacc agtggtgcag ggtcctccgg cagtcgttac ctgaagaaaa
     7021 aattccatca caaacatgca tcgcgagaag acacctggga ccaggcccaa cacaacatac
     7081 acctagcagg cgtgaccggt ggatcagggg acaaatacta caaacagaag tactccagga
     7141 acgactggaa tggagaatca gaggagtaca acaggcggcc aaagagctgg gtgaagtcaa
     7201 tcgaggcatt tggagagagc tatatttccg agaagaccaa aggggagatt tctcagcctg
     7261 gggcggctat caacgagcac aagaacggct ctggggggaa caatcctcac caagggtcct
     7321 tagacctgga gattcgaagc gaaggaggaa acatttatga ctgttgcatt aaagcccaag
     7381 aaggaactct cgctatccct tgctgtggat ttcccttatg gctattttgg ggactagagg
     7441 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     7501 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     7561 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     7621 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     7681 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     7741 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7801 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7861 gttcgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7921 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7981 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     8041 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     8101 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     8161 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8221 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8281 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     8341 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8401 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     8701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     9001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     9061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     9121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     9181 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     9241 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     9301 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     9361 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     9421 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     9481 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     9541 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     9601 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     9661 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9721 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9781 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9841 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9901 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9961 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    10021 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    10081 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    10141 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    10201 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
