LOCUS       Exported                5947 bp ds-DNA     circular SYN 20-JUL-2019
DEFINITION  AAV expression of EGFP from TBG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.TBG.PI.eGFP.WPRE.bGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5947)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 21, 2020 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5947
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    49..70
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        85..115
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1:  85 ..  90 / #ffffff / -35
                        2:  91 .. 108 / #ffffff
                        3: 109 .. 115 / #ffffff / -10"
     protein_bind    123..139
                     /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)."
                     /note="color: #31849b"
     primer_bind     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     147..163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     147..163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     repeat_region   201..330
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     primer_bind     complement(212..223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     273..284
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     promoter        670..1079
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
                     /note="color: #ffffff; direction: RIGHT"
     intron          1139..1271
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     primer_bind     1254..1273
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
                     /note="color: black; sequence: TTGCCTTTCTCTCCACAGGT"
     regulatory      1283..1287
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     regulatory      1288..1291
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1291..1332
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1960..1972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     CDS             2302..2328
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     primer_bind     complement(2314..2373)
                     /label=R199
                     /note="color: black; sequence: 
                     acaaattttgtaatccagaggttgattggatccaagcttttagtccagggtcaggcgct
                     c; added: 2019-07-17"
     primer_bind     complement(2400..2420)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(2818..2829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 2817"
                     /translation="IEGR"
     primer_bind     complement(2896..2913)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    2902..3109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3197..3326
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     primer_bind     complement(3243..3254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     3304..3315
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     complement(3345..3362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(3345..3361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(3354..3376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     rep_origin      3503..3958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(3590..3609)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     3800..3821
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     promoter        3984..4088
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4089..4949
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4089 .. 4157 / #ccffcc / signal sequence
                        2: 4158 .. 4949 / #ccffcc
                     Cleavage site after base 4157"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4307..4326)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      5120..5708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     5609..5628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     5862..5879
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
ORIGIN
        1 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
       61 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
      181 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      241 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      301 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      361 tatctacgta gccatgctct aggaagatcg gaattcgccc ttaagctagc aggttaattt
      421 ttaaaaagca gtcaaaagtc caagtggccc ttggcagcat ttactctctc tgtttgctct
      481 ggttaataat ctcaggagca caaacattcc agatccaggt taatttttaa aaagcagtca
      541 aaagtccaag tggcccttgg cagcatttac tctctctgtt tgctctggtt aataatctca
      601 ggagcacaaa cattccagat ccggcgcgcc agggctggaa gctacctttg acatcatttc
      661 ctctgcgaat gcatgtataa tttctacaga acctattaga aaggatcacc cagcctctgc
      721 ttttgtacaa ctttccctta aaaaactgcc aattccactg ctgtttggcc caatagtgag
      781 aactttttcc tgctgcctct tggtgctttt gcctatggcc cctattctgc ctgctgaaga
      841 cactcttgcc agcatggact taaacccctc cagctctgac aatcctcttt ctcttttgtt
      901 ttacatgaag ggtctggcag ccaaagcaat cactcaaagt tcaaacctta tcattttttg
      961 ctttgttcct cttggccttg gttttgtaca tcagctttga aaataccatc ccagggttaa
     1021 tgctggggtt aatttataac taagagtgct ctagttttgc aatacaggac atgctataaa
     1081 aatggaaaga tgttgctttc tgagagactg cagaagttgg tcgtgaggca ctgggcaggt
     1141 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1201 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1261 tctctccaca ggtgtccagg cggccgcatg ggcaagccca tccccaaccc cctgctgggc
     1321 ctggacagca ccgctagcaa agacaatact gtgcctctga agctgatcgc tctcctggct
     1381 aatggcgagt tccatagtgg cgaacagctg ggagaaaccc tgggcatgtc cagggccgct
     1441 atcaacaagc acattcagac tctgcgcgac tggggcgtgg acgtgttcac cgtgcccgga
     1501 aagggctact ctctgcccga gcctatcccg ctgctgaacg ctaaacagat tctgggacag
     1561 ctggacggcg ggagcgtggc agtcctgcct gtggtcgact ccaccaatca gtacctgctg
     1621 gatcgaatcg gcgagctgaa gagtggggat gcttgcattg cagaatatca gcaggcaggg
     1681 agaggaagca gagggaggaa atggttctct ccttttggag ctaacctgta cctgagtatg
     1741 ttttggcgcc tgaagcgggg accagcagca atcggcctgg gcccggtcat cggaattgtc
     1801 atggcagaag cgctgcgaaa gctgggagca gacaaggtgc gagtcaaatg gcccaatgac
     1861 ctgtatctgc aggatagaaa gctggcaggc atcctggtgg agctggccgg aataacaggc
     1921 gatgctgcac agatcgtcat tggcgccggg attaacgtgg ctatgaggcg cgtggaggaa
     1981 agcgtggtca atcagggctg gatcacactg caggaagcag ggattaacct ggacaggaat
     2041 actctggccg ctacgctgat ccgagagctg cgggcagccc tggaactgtt cgagcaggaa
     2101 ggcctggctc catatctgcc acggtgggag aagctggata acttcatcaa tagacccgtg
     2161 aagctgatca ttggggacaa agagattttc gggattagcc gggggattga taaacaggga
     2221 gccctgctgc tggaacagga cggagttatc aaaccctgga tgggcggaga aatcagtctg
     2281 cggtctgccg aaaagctgca gctgcctccc ctggagcgcc tgaccctgga ctaaaagctt
     2341 ggatccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2401 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2461 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2521 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2581 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2641 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2701 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2761 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2821 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2881 cgtcttcgag atctgcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2941 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3001 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3061 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggac tcgagttaag
     3121 ggcgaattcc cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     3181 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3241 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3301 gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta
     3361 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3421 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3481 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3661 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3841 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3901 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     3961 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4021 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4081 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4141 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4201 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4261 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4321 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4381 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4441 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4501 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4561 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4621 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4681 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4741 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4801 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4861 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4921 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4981 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5041 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5761 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5821 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5881 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5941 gcagctg
//
