LOCUS       Exported                5726 bp ds-DNA     circular SYN 11-DEC-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 5, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pOTTC1993 was constructed by Chris Richie at OTTC/NIDA (CR2679_E).
            
            
            backbone was pOTTC1464 was constructed by Chris Richie at OTTC/NIDA 
            (CR1689_H).
            Cre-independent iRFP-FLAG is flanked by NheI and AscI sites with the
            same linkers as are found in the DIO context (OTTC374 or CR1552_A).
            
            backbone = pOTTC374 was created by Chris Richie at OTTC/NIDA 
            (CR592_C, Addgene 47626).
FEATURES             Location/Qualifiers
     source          1..5726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1447..1812
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/locus_tag=source:synthetic DNA construct"
     LTR             1..141
                     /locus_tag="ITR (AAV)"
                     /label=ITR (AAV)
     LTR             1..117
                     /locus_tag="scITR (AAV, OTTC trs deletion)"
                     /label=scITR (AAV, OTTC trs deletion)
     promoter        156..1419
                     /locus_tag="EF-1a promoter"
                     /label=EF-1a promoter
     misc_recomb     1431..1440
                     /locus_tag="GEVVC NheI Infusion linker"
                     /label=GEVVC NheI Infusion linker
     misc_recomb     1431..1440
                     /locus_tag="GEVVC NheI Infusion linker(1)"
                     /label=GEVVC NheI Infusion linker
                     /label=GEVVC NheI Infusion linker(1)
     misc_recomb     1431..1439
                     /locus_tag="lox2272"
                     /label=lox2272
     misc_recomb     1441..1446
                     /locus_tag="GEVVC NheI Infusion linker(2)"
                     /label=GEVVC NheI Infusion linker
                     /label=GEVVC NheI Infusion linker(2)
     promoter        1447..1452
                     /locus_tag="EF-1-alpha promoter"
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human
                     elongationfactor EF-1-alpha"
     CDS             1483..1812
                     /codon_start=1
                     /locus_tag="CoV-2-E-2xStrep"
                     /label=CoV-2-E-2xStrep
                     /translation="MYSFVSEETGTLIVNSVLLFLAFVVFLLVTLAILTALRLCAYCCN
                     IVNVSLVKPSFYVYSRVKNLNSSRVPDLLVLEGGGGWSHPQFEKGGGSGGGSGGGSWSH
                     PQFEK"
     CDS             1726..1749
                     /codon_start=1
                     /locus_tag="Strep-Tag II"
                     /product="peptide that binds Strep-Tactin(R), an
                     engineeredform of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1786..1809
                     /codon_start=1
                     /locus_tag="Strep-Tag II(1)"
                     /product="peptide that binds Strep-Tactin(R), an
                     engineeredform of streptavidin"
                     /label=Strep-Tag II
                     /label=Strep-Tag II(1)
                     /translation="WSHPQFEK"
     misc_recomb     complement(1821..1829)
                     /locus_tag="GEVVC AscI Infusion linker"
                     /label=GEVVC AscI Infusion linker
     misc_recomb     complement(1821..1829)
                     /locus_tag="GEVVC AscI Infusion linker(1)"
                     /label=GEVVC AscI Infusion linker
                     /label=GEVVC AscI Infusion linker(1)
     misc_recomb     complement(1821..1828)
                     /locus_tag="loxP"
                     /label=loxP
     misc_RNA        1843..2451
                     /locus_tag="WPRE"
                     /label=WPRE
     polyA_signal    2472..2952
                     /locus_tag="hGH polyA"
                     /label=hGH polyA
     LTR             complement(2989..3129)
                     /locus_tag="ITR (AAV)(1)"
                     /label=ITR (AAV)
                     /label=ITR (AAV)(1)
     LTR             complement(3013..3129)
                     /locus_tag="scITR (AAV, OTTC trs deletion)(1)"
                     /label=scITR (AAV, OTTC trs deletion)
                     /label=scITR (AAV, OTTC trs deletion)(1)
     rep_origin      3204..3659
                     /locus_tag="M13 origin"
                     /label=M13 origin
     rep_origin      3221..3527
                     /locus_tag="f1 origin"
                     /label=f1 origin
     promoter        3947..4045
                     /locus_tag="Bla promoter (prokaryotic)"
                     /label=Bla promoter (prokaryotic)
     CDS             4046..4903
                     /locus_tag="AmpR"
                     /label=AmpR
     rep_origin      5001..5683
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgaagttatc
     1441 gctagctcgt gaggatctat ttccggtgaa ttcgccgcca ccatgtacag cttcgtatca
     1501 gaagaaaccg ggacactgat cgtaaattct gtgctcttgt ttctggcatt cgtcgtattt
     1561 ctcctcgtca cactggcaat tctgactgca ttgaggcttt gcgcctactg ttgtaacatt
     1621 gtcaatgtat ctctcgtgaa accctcattc tacgtttaca gcagggtgaa gaatctcaat
     1681 tctagcaggg tgccggatct cctcgttctc gaaggcggcg ggggatggag ccatccacaa
     1741 ttcgagaaag gcggtggttc aggaggaggt agcgggggtg gatcatggtc acatccgcag
     1801 tttgaaaagt aaggcgcgcc ataacttcga attcgatatc aagcttatcg ataatcaacc
     1861 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1921 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1981 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2041 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2101 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2161 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2221 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     2281 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2341 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2401 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     2461 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     2521 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     2581 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2641 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     2701 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     2761 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     2821 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2881 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     2941 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     3001 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3061 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3121 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3181 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3241 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3301 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3361 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3421 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3481 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3541 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3601 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3661 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3721 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3781 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3841 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3901 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3961 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4021 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4081 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4141 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4201 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4261 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4321 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4381 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4441 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4501 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4561 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4621 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4681 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5221 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5701 cttttgctgg ccttttgctc acatgt
//
