LOCUS       sequence-329517-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  AUTHORS   Berbari NF, Johnson AD, Lewis JS, Askwith CC, Mykytyn K
  TITLE     Identification of ciliary localization sequences within the third 
            intracellular loop of G protein-coupled receptors.
  JOURNAL   Mol Biol Cell. 2008 Apr;19(4):1540-7. Epub 2008 Feb 6.
  PUBMED    18256283
REFERENCE   2  (bases 1 to 5987)
  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: "Mol Biol 
            Cell."; date: "2008-04"; volume: "19(4)"; pages: "1540-7. Epub 2008 
            Feb 6"
COMMENT     Sequence Label: pEGFPN3-Sstr3
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1868..1877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1874..1876,1877..1879,1880..2593)
                     /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(1874..1876,1877..1879,1880..2590)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1919..1940)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2180..2199)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2527..2548
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2716..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2753..2772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2807..2826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2844..3299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2981..3002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3193..3212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3326..3430
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3432..3789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3432..3623
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3455..3475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3640..3775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3824..4618
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3824..4618
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3878..3897)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4488..4507
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4806..4825)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4850..4897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5226..5814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5715..5734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctcgaga tggccactgt tacctatccc tcatccgagc ctatgacctt
      601 ggaccctggg aacacatcct cgacctggcc cctggatacc accctgggga acacatccgc
      661 tggcgctagc ctgacaggcc tggctgtcag tggcatcttg atctctctgg tgtacctggt
      721 ggtgtgcgtg gtgggtctgc tgggcaactc gctggtgatc tacgtggtcc tgcggcacac
      781 gtccagccca tcagtgacca gtgtctatat cctcaacctg gctctggctg atgagctctt
      841 catgctaggg ctacccttcc tggctgctca gaacgccctg tcctactggc cctttggatc
      901 tctcatgtgc cgtctggtca tggccgtgga tggcatcaac cagttcacca gcatcttctg
      961 cctcaccgtc atgagtgtgg accgctatct ggctgtggtg caccccacac gctcagcccg
     1021 ctggcgcacg gcaccagtgg ctcgcacggt cagtgcagct gtctgggtgg cctcggctgt
     1081 ggtggtgctg cctgtggttg tgttctcagg agtgccccgg ggcatgagca cgtgccacat
     1141 gcagtggcca gagccagcgg ctgcctggcg aacagccttt atcatctaca cggccgcact
     1201 gggcttcttt gggcccctgc tggtcatctg cttgtgctac ttgctcattg tggtaaaggt
     1261 gcggtcgacc acccggcggg tgcgggcgcc ctcgtgtcag tgggtacagg cacccgcatg
     1321 ccagcggcga cgccgctctg agcgcagggt cacacgcatg gtggtggccg tggtggcact
     1381 cttcgtcctc tgctggatgc ctttctatct gctcaacatc gtcaatgtgg tgtgcccgct
     1441 gccggaggag cctgccttct tcggcctcta cttcctggtg gtggcgctgc cctatgccaa
     1501 cagctgcgca aaccccatcc tctacggctt cctctcctac cgcttcaagc agggctttcg
     1561 caggatcctg ctaagaccat cacgtcgcat tcggagccag gagccagggt cgggacctcc
     1621 agagaagact gaagaggagg aggatgaaga agaagaagag agaagggaag aggaggagcg
     1681 gaggatgcag agagggcagg agatgaacgg gaggctcagt cagatcgcac aggctggcac
     1741 tagtggacaa cagccacggc cctgcacagg gactgctaag gagcagcagc ttctgcccca
     1801 ggaggccaca gctggggaca aggccagcac actgagccat ctgggtaccg cgggcccggg
     1861 atccatcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1921 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1981 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2041 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2101 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2161 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2221 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2281 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2341 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2401 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2461 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2521 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2581 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     2641 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2701 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2761 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2821 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2881 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2941 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3001 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3061 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3121 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3181 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3241 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3301 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3361 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3421 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3721 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3781 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3841 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3901 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3961 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4021 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4081 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4141 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4201 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4261 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4321 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4381 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4441 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4501 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4561 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4621 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4681 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4741 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4801 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4861 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5041 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5881 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     5941 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttcc
//
