LOCUS       sequence-68604-e        5825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for homologous recombination that expresses ECFP 
            consitutively using the Aedes aegypti polyubiquitin (PUb) promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  TITLE     Vosshall lab Aedes aegypti HR donor plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5825)
  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            "Unpublished"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: pSL1180-HR-PUbECFP
FEATURES             Location/Qualifiers
     source          1..5825
                     /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"
     polyA_signal    289..423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(313..332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     367..386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(564..584)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(594..1304,1305..1307,1308..1310))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(636..657)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     985..1004
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1244..1265
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2953..2970)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2953..2969)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2962..2984)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3182..3637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3269..3288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3479..3500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3654..3673)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3773..3795
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3833..3851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3919..4023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4024..4092,4093..4884)
                     /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(4242..4261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5055..5643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5544..5563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5797..5814
                     /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 tccagacatg
      301 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      361 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      421 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
      481 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gttatctaga
      541 tccggtggat cttacgggtc ctccaccttc cgctttttct tgggtcgaga tctcttgtac
      601 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
      661 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
      721 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
      781 acgctgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc
      841 ttgtcggcgg tgatatagac gttgtggctg atgtagttgt actccagctt gtgccccagg
      901 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
      961 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     1021 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     1081 tcggggtagc ggctgaagca ctgcacgccc caggtcaggg tggtcacgag ggtgggccag
     1141 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     1201 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     1261 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gctagcgttg
     1321 aaatctctgt tgagcagaaa aagaaacgag gaaacgcttg agtaattggt tgtgaaatgc
     1381 aaactctcat ttgatattga ttcattgcct ttggcttcga gcacgacacg acaggtttta
     1441 aacttgtttt gctttgtctg cgtttgcagt cgcaggccaa gtgaaaaata tacacttgaa
     1501 ggtgatgacg tcacaacaac gccctacttt taagtgaaaa attaacttgt tttcgacttt
     1561 gaactacgta gttttgaaat tgcgtatctt caagttttca cgatttcctc aaggtttttc
     1621 tcgatatgtg ttaatattac cttaatgggt aattaccatc aaaatattta ttttagatat
     1681 gtgacggagc aaatacgtta ttcttattat tctagaaatt taattcaatt agtagcgatg
     1741 attcaacgaa atatgattat cgctgtgaat cacaattggg ttttatcaat gatgatgaaa
     1801 ctgcgttgca aattttcact aatcactcaa agctcaatag tcgccatctt ggaaaatagt
     1861 ttgctcattc aaagacaaag gaatcattca ccaaactagt tttcgctcat agctataatt
     1921 tcatcaatta atttacctac cttcactaga agattccctt caccgaaatg cactttcacc
     1981 aatttaatag taattgtcct ttgaataaag ctttgttcac tctgaaattt tctcctctgg
     2041 ctaattggat cactcttttt cactagagac ttcacttcac ttgcactggc actgcttact
     2101 tgggccgcgt aatgttcact ccactaggaa acgtattcga ttgagctggt ttcgcctttg
     2161 caggggcgtt ttatagacac tgtcgtagtg gtggttgtac ttctagaaaa tttcagcaat
     2221 acattcatac atatacctct gttcggttgg atggctctat atcgatcgaa tatgggtacc
     2281 atccctgtgt ctgaatggac agcaaaacgt gcttgtgtct gttagtcgtt cattctgtac
     2341 cttgaacgat gcagttcaac ttctggcaaa gacgtcaatg tacctaccct tcgtgtatat
     2401 ggcatagaga ggaagatgtg cgaatgcctt tttcgataga gaaaggattt cctgatttga
     2461 tcgacaattt cggtagctaa tcttagcttc gaatgtaatc tgataaccaa atccagagaa
     2521 aaatcagtaa tttgggaatt tcacttgaat cattctaatt gcatcatttg ctgtattagt
     2581 gcagtcagca agtgacgtca acccttctaa atcgatatac ttctgggaag ctttctttct
     2641 tgtctggctc agctggtgcc aaggcaaatt ataattggat tcaatgcaca agctacatgt
     2701 aaagatagct agcatgcgca aatttaaagc gctgatatca gcgctttaaa tttgcgcatg
     2761 ctagctatag ttctagaggt accggttgtt aacgttagcc ggctacgtat actccggaat
     2821 attaataggc ctaggatgca tatggcggcc gcctgcagct ggcgccatcg atacgcgtac
     2881 gtcgcgaccg cggacatgta cagagctcga gaagtactag tggccacgtg ggccgtgcac
     2941 cttaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     3121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3181 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3361 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3481 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     3541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3601 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3661 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3721 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3781 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3841 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3901 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3961 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4021 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4081 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4141 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4201 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4261 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4321 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4381 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4441 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4501 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4561 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4621 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4681 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4741 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4801 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4861 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5161 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5761 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5821 gaaga
//
