LOCUS       sequence-346308-        7116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transgenic LacZ reporter construct for validation of enhancer 
            sequences (inserted via Gibson-cloning). Can be introduced into the 
            mouse genome via random integration following pro-nuclear 
            injection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7116)
  TITLE     Hsp68-LacZ-Gibson plasmid for random transgenesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7116)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Hsp68-LacZ-Gibson
FEATURES             Location/Qualifiers
     source          1..7116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        100..118
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1027..4074
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     polyA_signal    4222..4356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4313..4332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4353..4369)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4353..4369)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4404..4423)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4405..4423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4430..4447)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4430..4446)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4587..5042)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4724..4745)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4936..4955
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5069..5173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5174..5242,5243..6034)
                     /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(5392..5411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6205..6793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6694..6713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6947..6964
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7081..7102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga
      121 acaaaagctg gtaccgagct ccaggaacat ccaaactgag cagccggggt cccccccacc
      181 ccccaccccg cccctcccgg caactttgag cctgtgctgg gacagagcct ctagttccta
      241 aattagtcca tgaggtcaga ggcagcactg ccattgtaac gcgattggag aggatcacgt
      301 caccggacac gcccccaggc atctccctgg gtctcctaaa cttggcgggg agaagtttta
      361 gcccttaagt tttagccttt aacccccata ttcagaactg tgcgagttgg cgaaacccca
      421 caaatcacaa caaactgtac acaacaccga gctagaggtg atctttcttg tccattccac
      481 acaggcctta gtaatgcgtc gccatagcaa cagtgtcact agtagcacca gcacttcccc
      541 acaccctccc cctcaggaat ccgtactctc cagtgaaccc cagaaacctc tggagagttc
      601 tggacaaggg cggaacccac aactccgatt actcaaggga ggcggggaag ctccaccaga
      661 cgcgaaactg ctggaagatt cctggcccca aggcctcctc cggctcgctg attggcccag
      721 cggagagtgg gcggggccgg tgaagactcc ttaaaggcgc agggcggcga gcaggtcacc
      781 agacgctgac agctactcag aaccaaatct ggttccatcc agagacaagc gaagacaaga
      841 gaagcagagc gagcggcgcg ttcccgatcc tcggccagga ccagccttcc ccagagcatc
      901 cctgccgcgg agcgcaacct tcccaggagc atccctgccg cggagcgcaa ctttccccgg
      961 agcatccacg ccgcggagcg cagccttcca gaagcagagc gcggcgccat ggctcgcgat
     1021 gatcccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1081 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     1141 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     1201 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     1261 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
     1321 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     1381 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     1441 aactcggcgt ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca ggacagtcgt
     1501 ttgccgtctg aatttgacct gagcgcattt ttacgcgccg gagaaaaccg cctcgcggtg
     1561 atggtgctgc gctggagtga cggcagttat ctggaagatc aggatatgtg gcggatgagc
     1621 ggcattttcc gtgacgtctc gttgctgcat aaaccgacta cacaaatcag cgatttccat
     1681 gttgccactc gctttaatga tgatttcagc cgcgctgtac tggaggctga agttcagatg
     1741 tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg tgaaacgcag
     1801 gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg atgagcgtgg tggttatgcc
     1861 gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc cgaaatcccg
     1921 aatctctatc gtgcggtggt tgaactgcac accgccgacg gcacgctgat tgaagcagaa
     1981 gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct gctgaacggc
     2041 aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca tggtcaggtc
     2101 atggatgagc agacgatggt gcaggatatc ctgctgatga agcagaacaa ctttaacgcc
     2161 gtgcgctgtt cgcattatcc gaaccatccg ctgtggtaca cgctgtgcga ccgctacggc
     2221 ctgtatgtgg tggatgaagc caatattgaa acccacggca tggtgccaat gaatcgtctg
     2281 accgatgatc cgcgctggct accggcgatg agcgaacgcg taacgcgaat ggtgcagcgc
     2341 gatcgtaatc acccgagtgt gatcatctgg tcgctgggga atgaatcagg ccacggcgct
     2401 aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc cttcccgccc ggtgcagtat
     2461 gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta cgcgcgcgtg
     2521 gatgaagacc agcccttccc ggctgtgccg aaatggtcca tcaaaaaatg gctttcgcta
     2581 cctggagaga cgcgcccgct gatcctttgc gaatacgccc acgcgatggg taacagtctt
     2641 ggcggtttcg ctaaatactg gcaggcgttt cgtcagtatc cccgtttaca gggcggcttc
     2701 gtctgggact gggtggatca gtcgctgatt aaatatgatg aaaacggcaa cccgtggtcg
     2761 gcttacggcg gtgattttgg cgatacgccg aacgatcgcc agttctgtat gaacggtctg
     2821 gtctttgccg accgcacgcc gcatccagcg ctgacggaag caaaacacca gcagcagttt
     2881 ttccagttcc gtttatccgg gcaaaccatc gaagtgacca gcgaatacct gttccgtcat
     2941 agcgataacg agctcctgca ctggatggtg gcgctggatg gtaagccgct ggcaagcggt
     3001 gaagtgcctc tggatgtcgc tccacaaggt aaacagttga ttgaactgcc tgaactaccg
     3061 cagccggaga gcgccgggca actctggctc acagtacgcg tagtgcaacc gaacgcgacc
     3121 gcatggtcag aagccgggca catcagcgcc tggcagcagt ggcgtctggc ggaaaacctc
     3181 agtgtgacgc tccccgccgc gtcccacgcc atcccgcatc tgaccaccag cgaaatggat
     3241 ttttgcatcg agctgggtaa taagcgttgg caatttaacc gccagtcagg ctttctttca
     3301 cagatgtgga ttggcgataa aaaacaactg ctgacgccgc tgcgcgatca gttcacccgt
     3361 gcaccgctgg ataacgacat tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg
     3421 gtcgaacgct ggaaggcggc gggccattac caggccgaag cagcgttgtt gcagtgcacg
     3481 gcagatacac ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca gcatcagggg
     3541 aaaaccttat ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt
     3601 accgttgatg ttgaagtggc gagcgataca ccgcatccgg cgcggattgg cctgaactgc
     3661 cagctggcgc aggtagcaga gcgggtaaac tggctcggat tagggccgca agaaaactat
     3721 cccgaccgcc ttactgccgc ctgttttgac cgctgggatc tgccattgtc agacatgtat
     3781 accccgtacg tcttcccgag cgaaaacggt ctgcgctgcg ggacgcgcga attgaattat
     3841 ggcccacacc agtggcgcgg cgacttccag ttcaacatca gccgctacag tcaacagcaa
     3901 ctgatggaaa ccagccatcg ccatctgctg cacgcggaag aaggcacatg gctgaatatc
     3961 gacggtttcc atatggggat tggtggcgac gactcctgga gcccgtcagt atcggcggaa
     4021 ttccagctga gcgccggtcg ctaccattac cagttggtct ggtgtcaaaa ataataataa
     4081 ccgggcaggg gggatctaag ctctagataa gtaatgatca taatcagcca tatcacatct
     4141 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4201 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4321 tccaaactca tcaatgtatc ttatcatgtc tggatccact agttctagag cggccgccac
     4381 cgcggtgcgt cgacgccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     4441 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4501 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4561 agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc
     4621 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4681 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4741 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4801 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4861 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4921 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4981 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5041 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5101 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5161 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5221 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5281 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5341 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5401 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5461 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5521 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5581 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5641 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5701 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5761 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5821 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5881 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5941 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6001 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6841 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6901 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6961 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7021 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     7081 taatgtgagt tagctcactc attaggcacc ccaggc
//
