LOCUS       Exported                5545 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector expressing Cre from TBG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV.TBG.PI.Cre.rBG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  TITLE     AAV plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5545
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        480..889
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          1027..1159
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1142..1161
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1204..1222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1273..2304
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(2382..2401)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2447..2502
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2595..2724
                     /label=AAV2 ITR
     primer_bind     complement(2743..2760)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2743..2759)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2752..2774)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2901..3356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3198..3219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3382..3486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3487..4347
                     /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(3705..3724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4518..5106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5007..5026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5260..5277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5394..5415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5430..5460
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5468..5484
                     /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     5473..5495
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5492..5508
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5492..5508
                     /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 tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gaattcgccc ttaagctagc aggttaattt ttaaaaagca
      241 gtcaaaagtc caagtggccc ttggcagcat ttactctctc tgtttgctct ggttaataat
      301 ctcaggagca caaacattcc agatccaggt taatttttaa aaagcagtca aaagtccaag
      361 tggcccttgg cagcatttac tctctctgtt tgctctggtt aataatctca ggagcacaaa
      421 cattccagat ccggcgcgcc agggctggaa gctacctttg acatcatttc ctctgcgaat
      481 gcatgtataa tttctacaga acctattaga aaggatcacc cagcctctgc ttttgtacaa
      541 ctttccctta aaaaactgcc aattccactg ctgtttggcc caatagtgag aactttttcc
      601 tgctgcctct tggtgctttt gcctatggcc cctattctgc ctgctgaaga cactcttgcc
      661 agcatggact taaacccctc cagctctgac aatcctcttt ctcttttgtt ttacatgaag
      721 ggtctggcag ccaaagcaat cactcaaagt tcaaacctta tcattttttg ctttgttcct
      781 cttggccttg gttttgtaca tcagctttga aaataccatc ccagggttaa tgctggggtt
      841 aatttataac taagagtgct ctagttttgc aatacaggac atgctataaa aatggaaaga
      901 tgttgctttc tgagagacag ctttattgcg gtagtttatc acagttaaat tgctaacgca
      961 gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact
     1021 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1081 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1141 tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt
     1201 acttaatacg actcactata ggctagcctc gagaattgta cactttactt aaaaccatta
     1261 tctgagtgtg aaatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc
     1321 gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg
     1381 ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc
     1441 aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta
     1501 tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac
     1561 atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt
     1621 atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg
     1681 ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag
     1741 gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa
     1801 attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat
     1861 attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg
     1921 gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac
     1981 tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta
     2041 tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag
     2101 gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg
     2161 cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc
     2221 aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg
     2281 cgcctgctgg aagatggcga ttagccatta acgcggcgtg gtacctctag agtcgacccg
     2341 ggcggcctcg aggacggggt gaactacgcc tgaggatccg atctttttcc ctctgccaaa
     2401 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     2461 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag caattcgttg
     2521 atctgaattt cgaccaccca taatacccat taccctggta gataagtagc atggcgggtt
     2581 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2641 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2701 ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg tcgttttaca
     2761 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2821 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2881 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3241 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3481 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3541 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3601 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3661 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3721 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3781 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3841 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3901 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3961 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4021 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4081 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4141 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4201 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4261 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5161 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5221 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5281 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5341 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     5401 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5461 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     5521 agatttaatt aaggccttaa ttagg
//
