LOCUS       Exported                4954 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Express gene of interest under EF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AV0-BWS-2.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4954)
  TITLE     Oka Lab plasmids
REFERENCE   2  (bases 1 to 4954)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4954
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        45..1223
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          276..1214
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1171..1191
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    1276..1864
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1329..1349)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1747..1758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1885..1966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1922..1941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1976..1995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2019..2159
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2019..2148
                     /label=AAV2 ITR
     rep_origin      2234..2689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2321..2340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2531..2552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2706..2725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2825..2847
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2885..2903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2971..3075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3076..3936
                     /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     complement(3294..3313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4107..4695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4596..4615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4757..4886
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
       61 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
      121 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
      181 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
      241 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
      301 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
      361 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
      421 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
      481 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
      541 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
      601 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
      661 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
      721 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
      781 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
      841 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
      901 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
      961 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     1021 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     1081 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     1141 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     1201 ttcttccatt tcaggtgtcg tgaggaattg gatccgttta aacgcggccg cgaattcgtc
     1261 gacaattcta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1321 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1381 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     1441 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     1501 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     1561 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1621 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     1681 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     1741 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     1801 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     1861 ccgcatcgat aagtaagtga ctagaacttg tttattgcag cttataatgg ttacaaataa
     1921 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1981 ttgtccaaac tcatctgcac gcgtggcgcg ccggccgcag gaacccctag tgatggagtt
     2041 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2101 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     2161 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     2221 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2281 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     2581 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     2641 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2701 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2761 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2821 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2881 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2941 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3001 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3061 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3121 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3181 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3241 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3301 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3361 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3421 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3481 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3541 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3601 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3661 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3721 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3781 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3841 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3901 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3961 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4021 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4081 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4141 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4201 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4261 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4321 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4381 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4441 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4501 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4561 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4621 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4681 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4741 ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     4801 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     4861 ggccaactcc atcactaggg gttcctgcgg ccggcgcgcc ctgcagtttc ccaagctttg
     4921 caaagatgga taaagtttta aacagagagg aatc
//
