LOCUS       sequence-238604-        3975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making acrosin-GFP-GPI RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3975)
  AUTHORS   Winuthayanon W, Bernhardt ML, Padilla-Banks E, Myers PH, Edin ML, 
            Lih FB, Hewitt SC, Korach KS, Williams CJ
  TITLE     Oviductal estrogen receptor alpha signaling prevents 
            protease-mediated embryo death.
  JOURNAL   Elife. 2015 Dec 1;4:e10453. doi: 10.7554/eLife.10453.
  PUBMED    26623518
REFERENCE   2  (bases 1 to 3975)
  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: "2015-12-1"; volume: "4:e10453. doi"; pages: " 
            10.7554/eLife.10453"
COMMENT     Sequence Label: pIVT/Acrosin-GFP-GPI
FEATURES             Location/Qualifiers
     source          1..3975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(144..149,150..167,168..174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..198
                     /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     187..209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     206..222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     206..222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        246..264
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     246..263
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     276..295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        276..294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           337..379
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      410..419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(506..508,509..511,512..1219)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(551..572)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(812..831)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1159..1180
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     3'UTR           1398..1523
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1581..1598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1581..1597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1590..1612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1806..1825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1925..1947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1985..2003)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2071..2175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2176..2244,2245..3036)
                     /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(2394..2413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3207..3795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3696..3715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3949..3966
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
      241 caacgattta ggtgacacta tagaagagaa ggaattaata cgactcacta tagggagaga
      301 gagagaatta ccctcactaa agggaggaga agcttgcttg ttctttttgc agaagctcag
      361 aataaacgct caactttggc agatcaagct tgcgtgcctg caggtcgacg ccaccatggt
      421 agagatgctg ccaactgtcg ctgtgctggt cttggcagtg tccgtggttg ccaaggataa
      481 caccacgctg caggaattcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      541 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      601 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      661 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
      721 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      781 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      841 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      901 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      961 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1021 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1081 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1141 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1201 cggcatggat gaactatacc aactcgagaa tggcggcata agcctgctgg ttcagaacac
     1261 atcctggatg ctgctgctgc tgctttccct ctccctcctc caagccctgg acttcatttc
     1321 tctgtaatct agaggatccc cgggtaccga gctcgaattc gcccttctag tgactgacta
     1381 ggatctggtt accactaaac cagcctcaag aacacccgaa tggagtctct aagctacata
     1441 ataccaactt acactttaca aaatgttgtc ccccaaaatg tagccattcg tatctgctcc
     1501 taataaaaag aaagtttctt cacattctaa aaaaaaaaaa aaaaaaaaaa aaaaaaaccc
     1561 ccccccccaa gggcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1621 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1681 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     1741 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     1801 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     1861 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1921 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1981 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2041 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2101 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2161 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2221 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2281 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2341 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2401 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2461 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2521 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2581 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2641 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2701 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2761 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2821 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2881 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2941 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3001 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3061 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3121 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3181 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3241 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3301 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3361 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3421 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3481 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3541 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3601 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3661 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3721 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3781 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3841 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3901 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3961 agcgaggaag cggaa
//
