LOCUS       sequence-424392-        7472 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 7472)
  AUTHORS   Guo W, Urizar E, Kralikova M, Mobarec JC, Shi L, Filizola M, Javitch
            JA
  TITLE     Dopamine D2 receptors form higher order oligomers at physiological 
            expression levels.
  JOURNAL   EMBO J. 2008 Sep 3. 27(17):2293-304.
  PUBMED    18668123
REFERENCE   2  (bases 1 to 7472)
  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: "EMBO J. 
            2008 Sep 3. 27(17):2293-304."
COMMENT     Sequence Label: pcDNA-D2s-L-Venus
FEATURES             Location/Qualifiers
     source          1..7472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1983..2002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2174..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2802..2821
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2802..2820
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4220..4229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4226..4228,4229..4231,4232..4948)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKS"
     primer_bind     complement(4271..4292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4532..4551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4879..4900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4964..4982)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4965..4982)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5002..5019)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5008..5232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5278..5706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5365..5384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5575..5596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5715..5735)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5720..6049
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5900..6035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5962..5981
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6116..6910
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLGEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6170..6189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6780..6799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7084..7205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7121..7140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7175..7194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7254..7270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7254..7270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7267..7289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7278..7294
                     /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(7302..7308,7309..7326,7327..7332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcaactccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcaggac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     1981 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     2041 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     2101 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     2161 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggtgtgta cggtgggagg tctatataag cagagctctc
     2761 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     2821 gagacccaag cttggtacca tgaagacgat catcgccctg agctacatct tctgcctggt
     2881 gttcgccgac tacaaggacg atgatgacgc catggatcca ctgaatctgt cctggtatga
     2941 tgatgatctg gagaggcaga actggagccg gcccttcaac gggtcagacg ggaaggcgga
     3001 cagaccccac tacaactact atgccacact gctcaccctg ctcatcgctg tcatcgtctt
     3061 cggcaacgtg ctggtgtgca tggctgtgtc ccgcgagaag gcgctgcaga ccaccaccaa
     3121 ctacctgatc gtcagcctcg cagtggccga cctcctcgtc gccacactgg tcatgccctg
     3181 ggttgtctac ctggaggtgg taggtgagtg gaaattcagc aggattcact gtgacatctt
     3241 cgtcactctg gacgtcatga tgtgcacggc gagcatcctg aacttgtgtg ccatcagcat
     3301 cgacaggtac acagctgtgg ccatgcccat gctgtacaat acgcgctaca gctccaagcg
     3361 ccgggtcacc gtcatgatct ccatcgtctg ggtcctgtcc ttcaccatct cctgcccact
     3421 cctcttcgga ctcaataacg cagaccagaa cgagtgcatc attgccaacc cggccttcgt
     3481 ggtctactcc tccatcgtct ccttctacgt gcccttcatt gtcaccctgc tggtctacat
     3541 caagatctac attgtcctcc gcagacgccg caagcgagtc aacaccaaac gcagcagccg
     3601 agctttcagg gcccacctga gggctccact aaaggaggct gcccggcgag cccaggagct
     3661 ggagatggag atgctctcca gcaccagccc acccgagagg acccggtaca gccccatccc
     3721 acccagccac caccagctga ctctccccga cccgtcccac catggtctcc acagcactcc
     3781 cgacagcccc gccaaaccag agaagaatgg gcatgccaaa gaccacccca agattgccaa
     3841 gatctttgag atccagacca tgcccaatgg caaaacgcgt acctccctca agaccatgag
     3901 ccgtaggaag ctctcccagc agaaggagaa gaaagccact cagatgctcg ccattgttct
     3961 cggcgtgttc atcatctgct ggctgccctt cttcatcaca cacatcctga acatacactg
     4021 tgactgcaac atcccgcctg tcctgtacag cgccttcacg tggctgggct atgtcaacag
     4081 cgccgtgaac cccatcatct acaccacctt caacattgag ttccgcaagg ccttcctgaa
     4141 gatcctccac tgcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
     4201 cggtaccgcg ggcccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     4261 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4321 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     4381 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     4441 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4501 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4561 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4621 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4681 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     4741 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4801 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaaactga gcaaagaccc
     4861 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4921 cggcatggac gagctgtaca agtcctaatc tagagggccc tattctatag tgtcacctaa
     4981 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     5041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5161 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5221 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     5281 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5341 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5401 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5461 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5521 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5581 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatctataag
     5641 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5701 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5761 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5821 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5881 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5941 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     6001 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     6061 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     6121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     6301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg gcgaagagca
     6601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     6661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7081 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7141 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7201 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7321 ataaagtgta aagcctgggg tgcctaatga gtgggctaac tcacattaat tgcgttgcgc
     7381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7441 cgcgcgggga gaggcggttt gcgtattggg cg
//
