LOCUS       sequence-417438-        6005 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Overexpression of FLAG-S1PR1-R120A mutant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  TITLE     Constructs expressing FLAG-S1PR1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6005)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCAGGS-Flag-S1pr1-R120A
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1747..1756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1756..1779
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3027..3046)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3092..3147
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3146..3165)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3508..3524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3508..3524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3521..3543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3532..3548
                     /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(3556..3562,3563..3580,3581..3586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3601..3622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3681..3876
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3727..3862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3882..4016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3919..3938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3973..3992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4084..4101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4255..4843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4335..4354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5014..5805,5806..5874))
                     /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     5637..5656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5875..5979)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgakggggg sggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgagctcccg
     1741 ggtaccgcca ccatggacta caaggatgac gatgataagg ggcccaccag cgtcccgctg
     1801 gtcaaggccc accgcagctc ggtctctgac tacgtcaact atgatatcat cgtccggcat
     1861 tacaactaca cgggaaagct gaatatcagc gcggacaagg agaacagcat taaactgacc
     1921 tcggtggtgt tcattctcat ctgctgcttt atcatcctgg agaacatctt tgtcttgctg
     1981 accatttgga aaaccaagaa attccaccga cccatgtact attttattgg caatctggcc
     2041 ctctcagacc tgttggcagg agtagcctac acagctaacc tgctcttgtc tggggccacc
     2101 acctacaagc tcactcccgc ccagtggttt ctggcggaag ggagtatgtt tgtggccctg
     2161 tcagcctccg tgttcagtct cctcgccatc gccattgagc gctatatcac aatgctgaaa
     2221 atgaaactcc acaacgggag caataacttc cgcctcttcc tgctaatcag cgcctgctgg
     2281 gtcatctccc tcatcctggg tggcctgcct atcatgggct ggaactgcat cagtgcgctg
     2341 tccagctgct ccaccgtgct gccgctctac cacaagcact atatcctctt ctgcaccacg
     2401 gtcttcactc tgcttctgct ctccatcgtc attctgtact gcagaatcta ctccttggtc
     2461 aggactcgga gccgccgcct gacgttccgc aagaacattt ccaaggccag ccgcagctct
     2521 gagaagtcgc tggcgctgct caagaccgta attatcgtcc tgagcgtctt catcgcctgc
     2581 tgggcaccgc tcttcatcct gctcctgctg gatgtgggct gcaaggtgaa gacctgtgac
     2641 atcctcttca gagcggagta cttcctggtg ttagctgtgc tcaactccgg caccaacccc
     2701 atcatttaca ctctgaccaa caaggagatg cgtcgggcct tcatccggat catgtcctgc
     2761 tgcaagtgcc cgagcggaga ctctgctggc aaattcaagc gacccatcat cgccggcatg
     2821 gaattcagcc gcagcaaatc ggacaattcc tcccaccccc agaaagacga aggggacaac
     2881 ccagagacca ttatgtcttc tggaaacgtc aactcttctt cctagctcga gatcgaccgc
     2941 cgaccattat gtcttctgga aacgtcaact cttcttccta gctcgagatc gatggccaat
     3001 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     3061 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     3121 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3181 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     3241 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     3301 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     3361 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     3421 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3481 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3661 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3721 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3781 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3841 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     3901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6001 acctg
//
