LOCUS       sequence-361835-        3826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of AIDm-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3826)
  AUTHORS   Camlin NJ, Evans JP
  TITLE     Auxin-inducible protein degradation as a novel approach for protein 
            depletion and reverse genetic discoveries in mammalian 
            oocytesdagger.
  JOURNAL   Biol Reprod. 2019 Oct 25;101(4):704-718. doi: 10.1093/biolre/ioz113.
  PUBMED    31299080
REFERENCE   2  (bases 1 to 3826)
  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"; doi: "10"; journalName: 
            "Biol Reprod"; date: "2019-10-25- 25"; volume: "101"; issue: "4"; 
            pages: "704-718"
COMMENT     Sequence Label: pIVT-AIDm-EGFP
FEATURES             Location/Qualifiers
     source          1..3826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..41)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(24..40)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(33..55)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(249..268)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     368..390
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(428..446)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        514..618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(619..687,688..1479)
                     /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(837..856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1650..2238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2139..2158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2392..2409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2526..2547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2562..2567,2568..2585,2586..2592)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2600..2616
                     /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     2605..2627
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2624..2640
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2624..2640
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2664..2682
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2664..2681
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2694..2713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2694..2712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2755..2797
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             join(2984..2986,2987..3700)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2984..2986,2987..3697)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3026..3047)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3287..3306)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3634..3655
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cccccccccc caagggcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
       61 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      181 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
      241 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
      301 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      361 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
      421 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
      481 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      541 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      601 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      661 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      721 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      781 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      841 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      901 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      961 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1021 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1081 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1141 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1201 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1261 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1321 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1381 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1441 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1501 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1561 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1621 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1681 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1741 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1801 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1861 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1921 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1981 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2041 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2101 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2161 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2221 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2281 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2341 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2401 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2461 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2521 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     2581 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     2641 catgattacg ccaagctcca acgatttagg tgacactata gaagagaagg aattaatacg
     2701 actcactata gggagagaga gagaattacc ctcactaaag ggaggagaag cttgcttgtt
     2761 ctttttgcag aagctcagaa taaacgctca actttggcag atcaagcttg cgtgcctgca
     2821 ggtcgacatg gactctaaag agaagtcagc ttgtcctaag gaccctgcca agccacctgc
     2881 gaaggcccaa gtagtcggtt ggccacccgt gcgcagctat aggaagaacg tgatggtgtc
     2941 ctgtcagaag tctagcgggg ggcccgaggc tagcggcgga tctgtgagca agggcgagga
     3001 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3061 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3121 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3181 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3241 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3301 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3361 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3421 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3481 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3541 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     3601 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3661 cgccgggatc actctcggca tggacgagct gtacaagtag tctagaggat ccccgggtac
     3721 cgagctcgaa ttcgcccttc tagtgactga ctaggatctg gttaccacta aaccagcctc
     3781 aagaacaccc gaatggagtc tctaagctac ataataccaa cttaca
//
