LOCUS       sequence-73022-e        5574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for homologous recombination that expresses dsRED 
            constitutively using the Aedes aegypti polyubiquitin (PUb) 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  TITLE     Aedes homologous recombination donor plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5574)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PSL1180polyUBdsRED
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1687..1689,1690..1692,1693..2367)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1870..1890)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2088..2107
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2282..2305
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2488..2609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2525..2544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2579..2598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2702..2719)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2702..2718)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2711..2733)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2931..3386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3018..3037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3228..3249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3403..3422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3522..3544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3582..3600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3668..3772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3773..3841,3842..4633)
                     /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(3991..4010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4804..5392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5293..5312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5546..5563
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aattcgaatg
      241 gccatgggac gtcgacctga ggtaattata acccgggccc tatatatgga tcctatcttt
      301 acatgtagct tgtgcattga atccaattat aatttgcctt ggcaccagct gagccagaca
      361 agaaagaaag cttcccagaa gtatatcgat ttagaagggt tgacgtcact tgctgactgc
      421 actaatacag caaatgatgc aattagaatg attcaagtga aattcccaaa ttactgattt
      481 ttctctggat ttggttatca gattacattc gaagctaaga ttagctaccg aaattgtcga
      541 tcaaatcagg aaatcctttc tctatcgaaa aaggcattcg cacatcttcc tctctatgcc
      601 atatacacga agggtaggta cattgacgtc tttgccagaa gttgaactgc atcgttcaag
      661 gtacagaatg aacgactaac agacacaagc acgttttgct gtccattcag acacagggat
      721 ggtacccata ttcgatcgat atagagccat ccaaccgaac agaggtatat gtatgaatgt
      781 attgctgaaa ttttctagaa gtacaaccac cactacgaca gtgtctataa aacgcccctg
      841 caaaggcgaa accagctcaa tcgaatacgt ttcctagtgg agtgaacatt acgcggccca
      901 agtaagcagt gccagtgcaa gtgaagtgaa gtctctagtg aaaaagagtg atccaattag
      961 ccagaggaga aaatttcaga gtgaacaaag ctttattcaa aggacaatta ctattaaatt
     1021 ggtgaaagtg catttcggtg aagggaatct tctagtgaag gtaggtaaat taattgatga
     1081 aattatagct atgagcgaaa actagtttgg tgaatgattc ctttgtcttt gaatgagcaa
     1141 actattttcc aagatggcga ctattgagct ttgagtgatt agtgaaaatt tgcaacgcag
     1201 tttcatcatc attgataaaa cccaattgtg attcacagcg ataatcatat ttcgttgaat
     1261 catcgctact aattgaatta aatttctaga ataataagaa taacgtattt gctccgtcac
     1321 atatctaaaa taaatatttt gatggttaat tacccattaa ggtaatatta acacatatcg
     1381 agaaaaacct tgaggaaatc gtgaaaactt gaagatacgc aatttcaaaa ctacgtagtt
     1441 caaagtcgaa aacaagttaa tttttcactt aaaagtaggg cgttgttgtg acgtcatcac
     1501 cttcaagtgt atatttttca cttggcctgc gactgcaaac gcagacaaag caaaacaagt
     1561 ttaaaacctg tcgtgtcgtg ctcgaagcca aaggcaatga atcaatatca aatgagagtt
     1621 tgcatttcac aaccaattac tcaagcgttt cctcgtttct ttttctgctc aacagagatt
     1681 tcaaccatgg tgcgctcctc caagaacgtc atcaaggagt tcatgcgctt caaggtgcgc
     1741 atggagggca ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1801 tacgagggcc acaacaccgt gaagctgaag gtgaccaagg gcggccccct gcccttcgcc
     1861 tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc
     1921 gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1981 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggctgc
     2041 ttcatctaca aggtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag
     2101 aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg
     2161 aagggcgaga tccacaaggc cctgaagctg aaggacggcg gccactacct ggtggagttc
     2221 aagtccatct acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggactcc
     2281 aagctggaca tcacctccca caacgaggac tacaccatcg tggagcagta cgagcgcacc
     2341 gagggccgcc accacctgtt cctgtagcgg ccgcgactct agatcataat cagccatacc
     2401 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2461 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2581 ggtttgtcca aactcatcaa tgtatcttaa agcttatcga tacgcgtacg tcgcgaccgc
     2641 ggacatgtac agagctcgag aagtactagt ggccacgtgg gccgtgcacc ttaagcttgg
     2701 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2761 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2821 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2881 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2941 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3001 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3061 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3121 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3181 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3241 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     3301 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3361 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3421 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3481 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3541 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3601 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4741 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4921 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
