LOCUS       Exported                7412 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of LacZ from TBG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.TBG.PI.LacZ .bGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7412
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        470..879
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          939..1071
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1054..1073
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1089..4163
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(1109..1125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4171..4188)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4177..4384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4472..4601
                     /label=AAV2 ITR
     primer_bind     complement(4620..4636)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4778..5233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4865..4884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5075..5096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5260..5364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5365..6225
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5583..5602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6396..6984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6885..6904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7138..7155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7272..7293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7308..7338
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7346..7362
                     /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)."
     primer_bind     7351..7373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7370..7386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7370..7386
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctagc aggttaattt ttaaaaagca gtcaaaagtc
      241 caagtggccc ttggcagcat ttactctctc tgtttgctct ggttaataat ctcaggagca
      301 caaacattcc agatccaggt taatttttaa aaagcagtca aaagtccaag tggcccttgg
      361 cagcatttac tctctctgtt tgctctggtt aataatctca ggagcacaaa cattccagat
      421 ccggcgcgcc agggctggaa gctacctttg acatcatttc ctctgcgaat gcatgtataa
      481 tttctacaga acctattaga aaggatcacc cagcctctgc ttttgtacaa ctttccctta
      541 aaaaactgcc aattccactg ctgtttggcc caatagtgag aactttttcc tgctgcctct
      601 tggtgctttt gcctatggcc cctattctgc ctgctgaaga cactcttgcc agcatggact
      661 taaacccctc cagctctgac aatcctcttt ctcttttgtt ttacatgaag ggtctggcag
      721 ccaaagcaat cactcaaagt tcaaacctta tcattttttg ctttgttcct cttggccttg
      781 gttttgtaca tcagctttga aaataccatc ccagggttaa tgctggggtt aatttataac
      841 taagagtgct ctagttttgc aatacaggac atgctataaa aatggaaaga tgttgctttc
      901 tgagagactg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
      961 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1021 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccagg
     1081 cggccgccat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     1141 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1201 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1261 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     1321 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     1381 cgcccatcta caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg
     1441 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     1501 gccagacgcg aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc
     1561 gctgggtcgg ttacggccag gacagtcgtt tgccgtctga atttgacctg agcgcatttt
     1621 tacgcgccgg agaaaaccgc ctcgcggtga tggtgctgcg ttggagtgac ggcagttatc
     1681 tggaagatca ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata
     1741 aaccgactac acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttcagcc
     1801 gcgctgtact ggaggctgaa gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa
     1861 cagtttcttt atggcagggt gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg
     1921 aaattatcga tgagcgtggt ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa
     1981 acccgaaact gtggagcgcc gaaatcccga atctctatcg tgcggtggtt gaactgcaca
     2041 ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga
     2101 ttgaaaatgg tctgctgctg ctgaacggca agccgttgct gattcgaggc gttaaccgtc
     2161 acgagcatca tcctctgcat ggtcaggtca tggatgagca gacgatggtg caggatatcc
     2221 tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc
     2281 tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa
     2341 cccacggcat ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga
     2401 gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt
     2461 cgctggggaa tgaatcaggc cacggcgcta atcacgacgc gctgtatcgc tggatcaaat
     2521 ctgtcgatcc ttcccgcccg gtgcagtatg aaggcggcgg agccgacacc acggccaccg
     2581 atattatttg cccgatgtac gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga
     2641 aatggtccat caaaaaatgg ctttcgctac ctggagagac gcgcccgctg atcctttgcg
     2701 aatacgccca cgcgatgggt aacagtcttg gcggtttcgc taaatactgg caggcgtttc
     2761 gtcagtatcc ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta
     2821 aatatgatga aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga
     2881 acgatcgcca gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgc
     2941 tgacggaagc aaaacaccag cagcagtttt tccagttccg tttatccggg caaaccatcg
     3001 aagtgaccag cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg
     3061 cgctggatgg taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta
     3121 aacagttgat tgaactgcct gaactaccgc agccggagag cgccgggcaa ctctggctca
     3181 cagtacgcgt agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcct
     3241 ggcagcagtg gcgtctggcg gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca
     3301 tcccgcatct gaccaccagc gaaatggatt tttgcatcga gctgggtaat aagcgttggc
     3361 aatttaaccg ccagtcaggc tttctttcac agatgtggat tggcgataaa aaacaactgc
     3421 tgacgccgct gcgcgatcag ttcacccgtg caccgctgga taacgacatt ggcgtaagtg
     3481 aagcgacccg cattgaccct aacgcctggg tcgaacgctg gaaggcggcg ggccattacc
     3541 aggccgaagc agcgttgttg cagtgcacgg cagatacact tgctgatgcg gtgctgatta
     3601 cgaccgctca cgcgtggcag catcagggga aaaccttatt tatcagccgg aaaacctacc
     3661 ggattgatgg tagtggtcaa atggcgatta ccgttgatgt tgaagtggcg agcgatacac
     3721 cgcatccggc gcggattggc ctgaactgcc agctggcgca ggtagcagag cgggtaaact
     3781 ggctcggatt agggccgcaa gaaaactatc ccgaccgcct tactgccgcc tgttttgacc
     3841 gctgggatct gccattgtca gacatgtata ccccgtacgt cttcccgagc gaaaacggtc
     3901 tgcgctgcgg gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc gacttccagt
     3961 tcaacatcag ccgctacagt caacagcaac tgatggaaac cagccatcgc catctgctgc
     4021 acgcggaaga aggcacatgg ctgaatatcg acggtttcca tatggggatt ggtggcgacg
     4081 actcctggag cccgtcagta tcggcggaat tccagctgag cgccggtcgc taccattacc
     4141 agttggtctg gtgtcaaaaa taaggatctg cctcgactgt gccttctagt tgccagccat
     4201 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4261 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4321 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4381 gggactcgag ttaagggcga attcccgatt aggatcttcc tagagcatgg ctacgtagat
     4441 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4501 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4561 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca
     4621 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4681 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4741 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4921 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4981 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccccgata gacggttttt
     5041 cgccctttga cgctggagtt cacgttcctc aatagtggac tcttgttcca aactggaaca
     5101 acactcaacc ctatctcggt ctattctttt gatttataag ggatttttcc gatttcggcc
     5161 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5221 acgtttataa tttcaggtgg catctttcgg ggaaatgtgc gcggaacccc tatttgttta
     5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5341 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5401 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5461 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caatagtggt
     5521 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5581 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5641 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5701 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5761 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5821 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5881 aacgacgagc gtgacaccac gatgcctgta gtaatggtaa caacgttgcg caaactatta
     5941 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6001 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6061 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6121 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6181 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6241 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6301 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6361 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6541 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6781 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6961 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7021 ttttgctgcg gttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7081 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7141 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7201 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7261 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7321 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7381 tatgaccatg attacgccag atttaattaa gg
//
