LOCUS       sequence-353881-        5852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mouse kappa opioid receptor (GFP tagged) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  AUTHORS   Muratspahic E, Tomasevic N, Koehbach J, Duerrauer L, Hadzic S, 
            Castro J, Schober G, Sideromenos S, Clark RJ, Brierley SM, Craik DJ,
            Gruber CW
  TITLE     Design of a Stable Cyclic Peptide Analgesic Derived from Sunflower 
            Seeds that Targets the kappa-Opioid Receptor for the Treatment of 
            Chronic Abdominal Pain.
  JOURNAL   J Med Chem. 2021 Jul 8;64(13):9042-9055. doi: 
            10.1021/acs.jmedchem.1c00158. Epub 2021 Jun 23.
  PUBMED    34162205
REFERENCE   2  (bases 1 to 5852)
  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: 
            "10.1021/acs.jmedchem.1c00158"; journalName: "J Med Chem"; date: 
            "2021-07-8- 8"; volume: "64"; issue: "13"; pages: "9042-9055"
COMMENT     Sequence Label: pEGFP-mKOR
FEATURES             Location/Qualifiers
     source          1..5852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1792..1801
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1798..1800,1801..1803,1804..2517)
                     /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(1798..1800,1801..1803,1804..2514)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1843..1864)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2104..2123)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2451..2472
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2640..2761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2677..2696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2731..2750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2768..3223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2905..2926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3117..3136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3250..3354
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3356..3713
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3356..3547
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3379..3399)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3564..3699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3748..4542
                     /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             3748..4542
                     /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(3802..3821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4412..4431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4730..4749)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4774..4821
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5150..5738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5639..5658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct catggagtcc cccattcaga tcttccgagg agatccaggc
      661 cctacctgct ctcccagtgc ttgccttctc cccaacagca gctcttggtt ccccaactgg
      721 gcagaatccg acagtaatgg cagtgtgggc tcagaggatc agcagctgga gtccgcgcac
      781 atctctccgg ccatccctgt tatcatcacc gctgtctact ctgtggtatt tgtggtgggc
      841 ttagtgggca attctctggt catgtttgtc atcatccgat acacgaagat gaagaccgca
      901 accaacatct acatatttaa cctggctttg gcagatgctt tggttactac cactatgccc
      961 tttcagagtg ctgtctactt gatgaattct tggccttttg gagatgtgct atgcaagatt
     1021 gtcatttcca ttgactacta caacatgttt accagcatat tcaccttgac catgatgagt
     1081 gtggaccgct acattgctgt gtgccaccct gtgaaagctt tggacttccg aacacctttg
     1141 aaagcaaaga tcatcaacat ctgcatttgg ctcctggcat catctgttgg tatatcagcg
     1201 atagtccttg gaggcaccaa agtcagggaa gatgtggatg tcattgaatg ctccttgcag
     1261 tttcctgatg atgaatattc ctggtgggat ctcttcatga agatctgtgt cttcgtcttt
     1321 gcctttgtga tcccagtcct catcatcatt gtctgctaca ccctgatgat cctgcgcctg
     1381 aagagtgtcc ggctcctgtc tggctcccga gagaaggacc gaaatctccg ccgcatcacc
     1441 aagctggtgc tggtagtagt tgcagtcttc atcatctgtt ggacccccat tcacatcttt
     1501 atcctggtgg aggctctggg aagcacctcc cacagcacag ctgccctctc cagctattat
     1561 ttctgtattg ccttgggtta taccaacagc agcctgaatc ctgttctcta tgcctttctg
     1621 gatgaaaact tcaagcggtg ttttagggac ttctgcttcc ctattaagat gcgaatggag
     1681 cgccagagca ccaatagagt tagaaacaca gttcaggatc ctgcttccat gagagatgtg
     1741 ggagggatga ataagccagt aaaggtaccg cgggcccggg atccaccggt cgccaccatg
     1801 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1861 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1921 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1981 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2041 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2101 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2161 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2221 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2281 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2341 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2401 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2461 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     2521 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2581 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2641 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2701 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2761 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2821 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2881 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2941 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3001 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3061 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3121 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3181 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     3241 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3301 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     3361 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3601 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3661 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     3721 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3781 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3841 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3901 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     3961 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4021 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4081 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4141 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4201 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4261 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4321 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4381 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4441 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4501 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4561 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4621 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4681 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     4741 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4801 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4861 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4921 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4981 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5041 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5281 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5821 attctgtgga taaccgtatt accgccatgc at
//
