LOCUS       sequence-202368-        7422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of LacZ from TBG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7422)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV.TBG.PI.LacZ .bGH
FEATURES             Location/Qualifiers
     source          1..7422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..33
                     /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     22..44
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     41..57
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     41..57
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   95..224
                     /label=AAV2 ITR
     promoter        564..973
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          1033..1165
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1148..1167
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1183..4257
                     /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(1203..1219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4265..4282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4271..4478
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4566..4695
                     /label=AAV2 ITR
     primer_bind     complement(4714..4730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4872..5327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4959..4978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5169..5190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5353..5457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5458..5526,5527..6318)
                     /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(5676..5695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6489..7077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6978..6997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7365..7386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7401..7406,7407..7422,1..2,3..9)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
       61 gattacgcca gatttaatta aggccttaat taggctgcgc gctcgctcgc tcactgaggc
      121 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      181 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      241 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcggaattc gcccttaagc
      301 tagcaggtta atttttaaaa agcagtcaaa agtccaagtg gcccttggca gcatttactc
      361 tctctgtttg ctctggttaa taatctcagg agcacaaaca ttccagatcc aggttaattt
      421 ttaaaaagca gtcaaaagtc caagtggccc ttggcagcat ttactctctc tgtttgctct
      481 ggttaataat ctcaggagca caaacattcc agatccggcg cgccagggct ggaagctacc
      541 tttgacatca tttcctctgc gaatgcatgt ataatttcta cagaacctat tagaaaggat
      601 cacccagcct ctgcttttgt acaactttcc cttaaaaaac tgccaattcc actgctgttt
      661 ggcccaatag tgagaacttt ttcctgctgc ctcttggtgc ttttgcctat ggcccctatt
      721 ctgcctgctg aagacactct tgccagcatg gacttaaacc cctccagctc tgacaatcct
      781 ctttctcttt tgttttacat gaagggtctg gcagccaaag caatcactca aagttcaaac
      841 cttatcattt tttgctttgt tcctcttggc cttggttttg tacatcagct ttgaaaatac
      901 catcccaggg ttaatgctgg ggttaattta taactaagag tgctctagtt ttgcaataca
      961 ggacatgcta taaaaatgga aagatgttgc tttctgagag actgcagaag ttggtcgtga
     1021 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     1081 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     1141 atccactttg cctttctctc cacaggtgtc caggcggccg ccatgaccat gattacggat
     1201 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1261 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1321 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     1381 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     1441 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     1501 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     1561 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     1621 gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg ccaggacagt
     1681 cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg
     1741 gtgatggtgc tgcgctggag tgacggcagt tatctggaag atcaggatat gtggcggatg
     1801 agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat cagcgatttc
     1861 catgttgcca ctcgctttaa tgatgatttc agccgcgctg tactggaggc tgaagttcag
     1921 atgtgcggcg agttgcgtga ctacctacgg gtaacagttt ctttatggca gggtgaaacg
     1981 caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg tggtggttat
     2041 gccgatcgcg tcacactacg tctgaacgtc gaaaacccga aactgtggag cgccgaaatc
     2101 ccgaatctct atcgtgcggt ggttgaactg cacaccgccg acggcacgct gattgaagca
     2161 gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa atggtctgct gctgctgaac
     2221 ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc atcatcctct gcatggtcag
     2281 gtcatggatg agcagacgat ggtgcaggat atcctgctga tgaagcagaa caactttaac
     2341 gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt acacgctgtg cgaccgctac
     2401 ggcctgtatg tggtggatga agccaatatt gaaacccacg gcatggtgcc aatgaatcgt
     2461 ctgaccgatg atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag
     2521 cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc
     2581 gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg cccggtgcag
     2641 tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat gtacgcgcgc
     2701 gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg
     2761 ctacctggag agacgcgccc gctgatcctt tgcgaatacg cccacgcgat gggtaacagt
     2821 cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt atccccgttt acagggcggc
     2881 ttcgtctggg actgggtgga tcagtcgctg attaaatatg atgaaaacgg caacccgtgg
     2941 tcggcttacg gcggtgattt tggcgatacg ccgaacgatc gccagttctg tatgaacggt
     3001 ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag
     3061 tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata cctgttccgt
     3121 catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc gctggcaagc
     3181 ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact gcctgaacta
     3241 ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca accgaacgcg
     3301 accgcatggt cagaagccgg gcacatcagc gcctggcagc agtggcgtct ggcggaaaac
     3361 ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg
     3421 gatttttgca tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt
     3481 tcacagatgt ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc
     3541 cgtgcaccgc tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc
     3601 tgggtcgaac gctggaaggc ggcgggccat taccaggccg aagcagcgtt gttgcagtgc
     3661 acggcagata cacttgctga tgcggtgctg attacgaccg ctcacgcgtg gcagcatcag
     3721 gggaaaacct tatttatcag ccggaaaacc taccggattg atggtagtgg tcaaatggcg
     3781 attaccgttg atgttgaagt ggcgagcgat acaccgcatc cggcgcggat tggcctgaac
     3841 tgccagctgg cgcaggtagc agagcgggta aactggctcg gattagggcc gcaagaaaac
     3901 tatcccgacc gccttactgc cgcctgtttt gaccgctggg atctgccatt gtcagacatg
     3961 tataccccgt acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat
     4021 tatggcccac accagtggcg cggcgacttc cagttcaaca tcagccgcta cagtcaacag
     4081 caactgatgg aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac atggctgaat
     4141 atcgacggtt tccatatggg gattggtggc gacgactcct ggagcccgtc agtatcggcg
     4201 gaattccagc tgagcgccgg tcgctaccat taccagttgg tctggtgtca aaaataagga
     4261 tctgcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4321 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4381 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4441 ggggaggatt gggaagacaa tagcaggcat gctggggact cgagttaagg gcgaattccc
     4501 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     4561 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4621 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4681 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
     4741 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4801 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4861 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4921 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4981 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5041 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5101 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5161 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5221 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5281 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5341 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5401 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5461 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5521 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5581 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5641 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5701 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5761 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5821 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5881 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5941 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6001 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6061 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6121 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6181 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6241 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6301 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6361 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6421 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7141 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7201 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7261 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7321 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7381 actcattagg caccccaggc tttacacttt atgcttccgg ct
//
