LOCUS       sequence-483454-        8129 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Constitutive expression of C5aR1-EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8129)
  AUTHORS   Suvorova ES, Gripentrog JM, Miettinen HM
  TITLE     Different endocytosis pathways of the C5a receptor and the N-formyl 
            peptide receptor.
  JOURNAL   Traffic. 2005 Feb;6(2):100-15.
  PUBMED    15634211
REFERENCE   2  (bases 1 to 8129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Traffic."; 
            date: "2005-02"; volume: "6(2)"; pages: "100-15"
COMMENT     Sequence Label: pBGSA C5aR1-EGFP
FEATURES             Location/Qualifiers
     source          1..8129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..269
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          309..405
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      1515..1524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1521..1523,1524..1526,1527..2240)
                     /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(1521..1523,1524..1526,1527..2237)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1566..1587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1827..1846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2174..2195
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2423..2442)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     promoter        complement(2771..2789)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2810..2826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2810..2826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2823..2845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2834..2850
                     /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(2858..2864,2865..2882,2883..2888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2903..2924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3041..3058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3212..3800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3292..3311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3971..4762,4763..4831))
                     /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     4594..4613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4832..4936)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4962..5417)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5099..5120)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5311..5330
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5544..5566
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5558..5575
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5559..5575
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5582..5601
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5582..5600
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5637..5771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5661..5680)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5715..5734
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6196..6216)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          6346..6411
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(6831..7625)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6942..6961)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7552..7571
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
       61 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
      121 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
      181 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
      241 tcgttttctg ttctgcgccg ttacagatcc aagctcctcg aggaactgaa aaaccagaaa
      301 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
      361 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
      421 ttacttctgc tctaaaagct gcggaattca tgaactcctt caattatacc acccctgatt
      481 atgggcacta tgatgacaag gataccctgg acctcaacac ccctgtggat aaaacttcta
      541 acacgctgcg tgttccagac atcctggcct tggtcatctt tgcagtcgtc ttcctggtgg
      601 gagtgctggg caatgccctg gtggtctggg tgacggcatt cgaggccaag cggaccatca
      661 atgccatctg gttcctcaac ttggcggtag ccgacttcct ctcctgcctg gcgctgccca
      721 tcttgttcac gtccattgta cagcatcacc actggccctt tggcggggcc gcctgcagca
      781 tcctgccctc cctcatcctg ctcaacatgt acgccagcat cctgctcctg gccaccatca
      841 gcgccgaccg ctttctgctg gtgtttaaac ccatctggtg ccagaacttc cgaggggccg
      901 gcttggcctg gatcgcctgt gccgtggctt ggggtttagc cctgctgctg accataccct
      961 ccttcctgta ccgggtggtc cgggaggagt actttccacc aaaggtgttg tgtggcgtgg
     1021 actacagcca cgacaaacgg cgggagcgag ccgtggccat cgtccggctg gtcctgggct
     1081 tcctgtggcc tctactcacg ctcacgattt gttacacttt catcctgctc cggacgtgga
     1141 gccgcagggc cacgcggtcc accaagacac tcaaggtggt ggtggcagtg gtggccagtt
     1201 tctttatctt ctggttgccc taccaggtga cggggataat gatgtccttc ctggagccat
     1261 cgtcacccac cttcctgctg ctgaataagc tggactccct gtgtgtctcc tttgcctaca
     1321 tcaactgctg catcaacccc atcatctacg tggtggccgg ccagggcttc cagggccgac
     1381 tgcggaaatc cctccccagc ctcctccgga acgtgttgac tgaagagtcc gtggttaggg
     1441 agagcaagtc attcacgcgc tccacagtgg acactatggc ccagaagacc caggcagtgc
     1501 gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1561 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1621 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1681 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1741 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1801 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1861 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1921 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1981 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2041 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2101 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2161 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2221 tggacgagct gtacaagtaa agcggccgcg actctagacc tggccttgga agttgccact
     2281 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     2341 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     2401 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     2461 tggctcactg caatctccgc ctcctgggtt caagcgattc ccctgcctca gcctcccgag
     2521 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     2581 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     2641 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     2701 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gcgagctcca
     2761 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     2821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2941 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4981 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     5041 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5101 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5161 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5221 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5281 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5341 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5401 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5461 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5521 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     5581 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg tcgaccgatc
     5641 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5701 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     5761 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     5821 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctct
     5881 agtcaaggca ctatacatca aatattcctt attaacccct ttacaaatta aaaagctaaa
     5941 ggtacacaat ttttgagcat agttattaat agcagacact ctatgcctgt gtggagtaag
     6001 aaaaaacagt atgttatgat tataactgtt atgcctactt ataaaggtta cagaatattt
     6061 ttccataatt ttcttgtata gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca
     6121 agcaagagtt ctattactaa acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa
     6181 gtccttgggg tcttctacct ttctcttctt ttttggagga gtagaatgtt gagagtcagc
     6241 agtagcctca tcatcactag atggcatttc ttctgagcaa aacaggtttt cctcattaaa
     6301 ggcattccac cactgctccc attcatcagt tccataggtt ggaatctaaa atacacaaac
     6361 aattagaatc agtagtttaa cacattatac acttaaaaat tttatattta ccttagagct
     6421 ttaaatctct gtaggtagtt tgtccaatta tgtcacacca cagaagtaag gttccttcac
     6481 aaagatccgg accaaagcgg ccatcgtgcc tccccactcc tgcagttcgg gggcatggat
     6541 gcgcggatag ccgctgctgg tttcctggat gccgacggat ttgcactgcc ggtagaactc
     6601 cgcgaggtcg tccagcctca ggcagcagct gaaccaactc gcgaggggat cgagcccggg
     6661 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     6721 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     6781 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     6841 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     6901 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     6961 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     7021 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     7081 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     7141 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     7201 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     7261 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     7321 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     7381 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     7441 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     7501 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     7561 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     7621 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     7681 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     7741 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     7801 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     7861 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     7921 tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgaagct
     7981 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     8041 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     8101 taggcttttg caaaaagctc ctcgatcga
//
