LOCUS       sequence-237438-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for in vivo enhancer reporter assay. Plasmid contains a 
            beta-globin minimal promoter followed by a lacZ reporter gene 
            derived from pRS16, with the entire reporter cassette flanked by 
            attB sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Castro JP, Yancoskie MN, Marchini M, Belohlavy S, Hiramatsu L, Kucka
            M, Beluch WH, Naumann R, Skuplik I, Cobb J, Barton NH, Rolian C, 
            Chan YF
  TITLE     An integrative genomic analysis of the Longshanks selection 
            experiment for longer limbs in mice.
  JOURNAL   Elife. 2019 Jun 6;8:e42014. doi: 10.7554/eLife.42014.
  PUBMED    31169497
REFERENCE   2  (bases 1 to 6685)
  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: "Elife."; 
            date: "2019-06-6"; volume: "8:e42014. doi"; pages: " 
            10.7554/eLife.42014"
COMMENT     Sequence Label: pBeta-lacZ-attBx2
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    669..738
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     763..779
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     764..780
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             922..3969
                     /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    4233..4367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4324..4343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4374..4390)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4374..4390)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    4391..4460
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(4497..4517)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4497..4515)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4536..4552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4536..4552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4549..4571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4560..4576
                     /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)."
     promoter        complement(join(4584..4590,4591..4608,4609..4614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4767..4784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4938..5526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5018..5037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5697..6488,6489..6557))
                     /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     6320..6339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6558..6662)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacatc
      661 gcggccgcct cgaagccgcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg
      721 tactccacct cacccatcgg taccgatgta ccgggccccc cctcgaggtc gacggtatcg
      781 ataagcttga tatcgaattc ctgcagcccg ggctgggcat aaaagtcagg gcagagccat
      841 ctattgctta catttgcttc tagcctgcag gtcgaggagc gcagccttcc agaagcagag
      901 cgcggcgcca tgggggatcc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      961 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1021 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc
     1081 tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc
     1141 gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc
     1201 aacgtaacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt
     1261 tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt
     1321 atttttgatg gcgttaactc ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac
     1381 ggccaggaca gtcgtttgcc gtctgaattt gacctgagcg catttttacg cgccggagaa
     1441 aaccgcctcg cggtgatggt gctgcgttgg agtgacggca gttatctgga agatcaggat
     1501 atgtggcgga tgagcggcat tttccgtgac gtctcgttgc tgcataaacc gactacacaa
     1561 atcagcgatt tccatgttgc cactcgcttt aatgatgatt tcagccgcgc tgtactggag
     1621 gctgaagttc agatgtgcgg cgagttgcgt gactacctac gggtaacagt ttctttatgg
     1681 cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg gcggtgaaat tatcgatgag
     1741 cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg tcgaaaaccc gaaactgtgg
     1801 agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac tgcacaccgc cgacggcacg
     1861 ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg tgcggattga aaatggtctg
     1921 ctgctgctga acggcaagcc gttgctgatt cgaggcgtta accgtcacga gcatcatcct
     1981 ctgcatggtc aggtcatgga tgagcagacg atggtgcagg atatcctgct gatgaagcag
     2041 aacaacttta acgccgtgcg ctgttcgcat tatccgaacc atccgctgtg gtacacgctg
     2101 tgcgaccgct acggcctgta tgtggtggat gaagccaata ttgaaaccca cggcatggtg
     2161 ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg
     2221 cgaatggtgc agcgcgatcg taatcacccg agtgtgatca tctggtcgct ggggaatgaa
     2281 tcaggccacg gcgctaatca cgacgcgctg tatcgctgga tcaaatctgt cgatccttcc
     2341 cgcccggtgc agtatgaagg cggcggagcc gacaccacgg ccaccgatat tatttgcccg
     2401 atgtacgcgc gcgtggatga agaccagccc ttcccggctg tgccgaaatg gtccatcaaa
     2461 aaatggcttt cgctacctgg agagacgcgc ccgctgatcc tttgcgaata cgcccacgcg
     2521 atgggtaaca gtcttggcgg tttcgctaaa tactggcagg cgtttcgtca gtatccccgt
     2581 ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc tgattaaata tgatgaaaac
     2641 ggcaacccgt ggtcggctta cggcggtgat tttggcgata cgccgaacga tcgccagttc
     2701 tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc cagcgctgac ggaagcaaaa
     2761 caccagcagc agtttttcca gttccgttta tccgggcaaa ccatcgaagt gaccagcgaa
     2821 tacctgttcc gtcatagcga taacgagctc ctgcactgga tggtggcgct ggatggtaag
     2881 ccgctggcaa gcggtgaagt gcctctggat gtcgctccac aaggtaaaca gttgattgaa
     2941 ctgcctgaac taccgcagcc ggagagcgcc gggcaactct ggctcacagt acgcgtagtg
     3001 caaccgaacg cgaccgcatg gtcagaagcc gggcacatca gcgcctggca gcagtggcgt
     3061 ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc acgccatccc gcatctgacc
     3121 accagcgaaa tggatttttg catcgagctg ggtaataagc gttggcaatt taaccgccag
     3181 tcaggctttc tttcacagat gtggattggc gataaaaaac aactgctgac gccgctgcgc
     3241 gatcagttca cccgtgcacc gctggataac gacattggcg taagtgaagc gacccgcatt
     3301 gaccctaacg cctgggtcga acgctggaag gcggcgggcc attaccaggc cgaagcagcg
     3361 ttgttgcagt gcacggcaga tacacttgct gatgcggtgc tgattacgac cgctcacgcg
     3421 tggcagcatc aggggaaaac cttatttatc agccggaaaa cctaccggat tgatggtagt
     3481 ggtcaaatgg cgattaccgt tgatgttgaa gtggcgagcg atacaccgca tccggcgcgg
     3541 attggcctga actgccagct ggcgcaggta gcagagcggg taaactggct cggattaggg
     3601 ccgcaagaaa actatcccga ccgccttact gccgcctgtt ttgaccgctg ggatctgcca
     3661 ttgtcagaca tgtatacccc gtacgtcttc ccgagcgaaa acggtctgcg ctgcgggacg
     3721 cgcgaattga attatggccc acaccagtgg cgcggcgact tccagttcaa catcagccgc
     3781 tacagtcaac agcaactgat ggaaaccagc catcgccatc tgctgcacgc ggaagaaggc
     3841 acatggctga atatcgacgg tttccatatg gggattggtg gcgacgactc ctggagcccg
     3901 tcagtatcgg cggaattcca gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt
     3961 caaaaataat aataaccggg caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt
     4021 atgtactatt taaaaaacac aaacttttgg atgttcggtt tattcttttt cttttacttt
     4081 tttatcatgg gagcctactt cccgtttttc ccgatttggc tacatgacat caaccatatc
     4141 agcaaaagtg atacgggtat tatttttgcc gctatttctc tgttctcgct attattccaa
     4201 ccgctgtttg gtctgctttc tgacaaactc ggaacttgtt tattgcagct tataatggtt
     4261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4321 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccg ggggatccac
     4381 tagttctaga ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg
     4441 cgtactccac ctcacccatc gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     4501 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4561 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4621 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4741 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4801 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5341 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5821 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6661 cgcacatttc cccgaaaagt gccac
//
