LOCUS       sequence-238658-        3060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making cRNA appropriate for microinjection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3060)
  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 3060)
  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: pIVT2
FEATURES             Location/Qualifiers
     source          1..3060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           8..50
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    57..113
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     3'UTR           151..276
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(337..354)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(337..353)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(346..368)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(562..581)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     681..703
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(741..759)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        827..931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(932..1000,1001..1792)
                     /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(1150..1169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1963..2551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2452..2471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2705..2722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2839..2860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2875..2880,2881..2898,2899..2905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2913..2929
                     /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     2918..2940
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2937..2953
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2937..2953
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2977..2995
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2977..2994
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3007..3026
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3007..3025
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttgctt gttctttttg cagaagctca gaataaacgc tcaactttgg cagatcaagc
       61 ttgcgtgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcgcccttc
      121 tagtgactga ctaggatctg gttaccacta aaccagcctc aagaacaccc gaatggagtc
      181 tctaagctac ataataccaa cttacacttt acaaaatgtt gtcccccaaa atgtagccat
      241 tcgtatctgc tcctaataaa aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa
      301 aaaaaaaaaa amcccccccc ccccaagggc gaattcactg gccgtcgttt tacaacgtcg
      361 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      421 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      481 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      541 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
      601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      721 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
      781 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      841 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      901 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      961 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1021 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1081 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1141 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1201 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1261 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1321 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1381 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1441 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1501 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1561 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1621 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     1681 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1741 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1801 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2101 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2641 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2701 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2761 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2821 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     2881 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     2941 aaacagctat gaccatgatt acgccaagct ccaacgattt aggtgacact atagaagaga
     3001 aggaattaat acgactcact atagggagag agagagaatt accctcacta aagggaggag
//
