LOCUS       sequence-345450-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The synthetic 3'UTR#5 for EGFP expression under the control of the 
            Ae. aegypti polyubiquitin (PUb) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  AUTHORS   Chae K, Valentin C, Jakes E, Myles KM, Adelman ZN
  TITLE     Novel synthetic 3'-untranslated regions for controlling transgene 
            expression in transgenic Aedes aegypti mosquitoes.
  JOURNAL   RNA Biol. 2021 Aug 31:1-9. doi: 10.1080/15476286.2021.1971440.
  PUBMED    34464234
REFERENCE   2  (bases 1 to 5630)
  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: "RNA Biol. 
            2021 Aug 31:1-9. doi: 10.1080/15476286.2021.1971440."
COMMENT     Sequence Label: pSLfa-PUb-EGFP-3'UTR#5
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2031..2053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2153..2172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2189..2644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2538..2557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2842..2864
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2856..2873
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2857..2873
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4374..4400
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4407..4409,4410..4412,4413..5126)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4407..4409,4410..4412,4413..5123)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4452..4473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4713..4732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5060..5081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5411..5427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5411..5427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5424..5446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5435..5451
                     /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(5459..5465,5466..5483,5484..5489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5504..5525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     2221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2281 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     2341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2401 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     2461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2641 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2701 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2761 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2821 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     2881 tggccggcct aggcgcgcca agcttaaggt gcacggccca cgtggccact agtacttctc
     2941 gagctctgta catgtccgcg gtcgcgacgt acgcgtatct ttacatgtag cttgtgcatt
     3001 gaatccaatt ataatttgcc ttggcaccag ctgagccaga caagaaagaa agcttcccag
     3061 aagtatatcg atttagaagg gttgacgtca cttgctgact gcactaatac agcaaatgat
     3121 gcaattagaa tgattcaagt gaaattccca aattactgat ttttctctgg atttggttat
     3181 cagattacat tcgaagctaa gattagctac cgaaattgtc gatcaaatca ggaaatcctt
     3241 tctctatcga aaaaggcatt cgcacatctt cctctctatg ccatatacac gaagggtagg
     3301 tacattgacg tctttgccag aagttgaact gcatcgttca aggtacagaa tgaacgacta
     3361 acagacacaa gcacgttttg ctgtccattc agacacaggg atggtaccca tattcgatcg
     3421 atatagagcc atccaaccga acagaggtat atgtatgaat gtattgctga aattttctag
     3481 aagtacaacc accactacga cagtgtctat aaaacgcccc tgcaaaggcg aaaccagctc
     3541 aatcgaatac gtttcctagt ggagtgaaca ttacgcggcc caagtaagca gtgccagtgc
     3601 aagtgaagtg aagtctctag tgaaaaagag tgatccaatt agccagagga gaaaatttca
     3661 gagtgaacaa agctttattc aaaggacaat tactattaaa ttggtgaaag tgcatttcgg
     3721 tgaagggaat cttctagtga aggtaggtaa attaattgat gaaattatag ctatgagcga
     3781 aaactagttt ggtgaatgat tcctttgtct ttgaatgagc aaactatttt ccaagatggc
     3841 gactattgag ctttgagtga ttagtgaaaa tttgcaacgc agtttcatca tcattgataa
     3901 aacccaattg tgattcacag cgataatcat atttcgttga atcatcgcta ctaattgaat
     3961 taaatttcta gaataataag aataacgtat ttgctccgtc acatatctaa aataaatatt
     4021 ttgatggtaa ttacccatta aggtaatatt aacacatatc gagaaaaacc ttgaggaaat
     4081 cgtgaaaact tgaagatacg caatttcaaa actacgtagt tcaaagtcga aaacaagtta
     4141 atttttcact taaaagtagg gcgttgttgt gacgtcatca ccttcaagtg tatatttttc
     4201 acttggcctg cgactgcaaa cgcagacaaa gcaaaacaag tttaaaacct gtcgtgtcgt
     4261 gctcgaagcc aaaggcaatg aatcaatatc aaatgagagt ttgcatttca caaccaatta
     4321 ctcaagcgtt tcctcgtttc tttttctgct caacagagat ttcaaccatg gagtacccat
     4381 acgacgtacc agattacgct catatgatgg tgagcaaggg cgaggagctg ttcaccgggg
     4441 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4501 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4561 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     4621 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4681 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4741 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4801 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4861 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4921 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4981 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     5041 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     5101 tcggcatgga cgagctgtac aagtaagtcg acgcggccgc cgaacgcaaa acatgggaca
     5161 attgtaaagt ggaagtccta aagtagtttt ccccgcaagc tctgctttac aaataaacat
     5221 tcaataaagt gctctcatca tattccgaga tattaacctt ttatgtacct attgtttttt
     5281 agatccatag tctcaatttc tatacatcca tcacaatcat tcaaacttat cattttttta
     5341 tatagaagca ctacatagtt cacgtgctcg agagaattcg gccggcctag gcgcgccaat
     5401 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5461 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5521 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
//
