LOCUS       sequence-59870-e        9435 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 9435)
  TITLE     BC-GPR56
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9435)
  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: 
            "Unpublished"
COMMENT     Sequence Label: BC-GPR56
FEATURES             Location/Qualifiers
     source          1..9435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      3214..3223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3656..3673
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3657..3673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3680..3699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3680..3698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3749..4210
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3805..3822)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4032..4051
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4251..4847
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4251..4270)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4747..4767
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5748..6263
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6432..6448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6432..6448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6445..6467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6456..6472
                     /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(6480..6486,6487..6504,6505..6510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6525..6546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6663..6680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6834..7422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6914..6933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7593..8384,8385..8453))
                     /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     8216..8235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8454..8558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8626..8644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8682..8704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8804..8823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9215..9234
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(9434..9435,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaaat tagatctacc ggtaagagtg actccgtcgg
     1441 aggaaaatga ctccccagtc gctgctgcag acgacactgt tcctgctgag tctgctcttc
     1501 ctggtccaag gtgcccacgg caggggccac agggaagact ttcgcttctg cagccagcgg
     1561 aaccagacac acaggagcag cctccactac aaacccacac cagacctgcg catctccatc
     1621 gagaactccg aagaggccct cacagtccat gcccctttcc ctgcagccca ccctgcttcc
     1681 cgatccttcc ctgaccccag gggcctctac cacttctgcc tctactggaa ccgacatgct
     1741 gggagattac atcttctcta tggcaagcgt gacttcttgc tgagtgacaa agcctctagc
     1801 ctcctctgct tccagcacca ggaggagagc ctggctcagg gccccccgct gttagccact
     1861 tctgtcacct cctggtggag ccctcagaac atcagcctgc ccagtgccgc cagcttcacc
     1921 ttctccttcc acagtcctcc ccacacggcc gctcacaatg cctcggtgga catgtgcgag
     1981 ctcaaaaggg acctccagct gctcagccag ttcctgaagc atccccagaa ggcctcaagg
     2041 aggccctcgg ctgcccccgc cagccagcag ttgcagagcc tggagtcgaa actgacctct
     2101 gtgagattca tgggggacat ggtgtccttc gaggaggacc ggatcaacgc cacggtgtgg
     2161 aagctccagc ccacagccgg cctccaggac ctgcacatcc actcccggca ggaggaggag
     2221 cagagcgaga tcatggagta ctcggtgctg ctgcctcgaa cactcttcca gaggacgaaa
     2281 ggccggagcg gggaggctga gaagagactc ctcctggtgg acttcagcag ccaagccctg
     2341 ttccaggaca agaattccag ccacgtcctg ggtgagaagg tcttggggat tgtggtacag
     2401 aacaccaaag tagccaacct cacggagccc gtggtgctca ccttccagca ccagctacag
     2461 ccgaagaatg tgactctgca atgtgtgttc tgggttgaag accccacatt gagcagcccg
     2521 gggcattgga gcagtgctgg gtgtgagacc gtcaggagag aaacccaaac atcctgcttc
     2581 tgcaaccact tgacctactt tgcagtgctg atggtctcct cggtggaggt ggacgccgtg
     2641 cacaagcact acctgagcct cctctcctac gtgggctgtg tcgtctctgc cctggcctgc
     2701 cttgtcacca ttgccgccta cctctgctcc agggtgcccc tgccgtgcag gaggaaacct
     2761 cgggactaca ccatcaaggt gcacatgaac ctgctgctgg ccgtcttcct gctggacacg
     2821 agcttcctgc tcagcgagcc ggtggccctg acaggctctg aggctggctg ccgagccagt
     2881 gccatcttcc tgcacttctc cctgctcacc tgcctttcct ggatgggcct cgaggggtac
     2941 aacctctacc gactcgtggt ggaggtcttt ggcacctatg tccctggcta cctactcaag
     3001 ctgagcgcca tgggctgggg cttccccatc tttctggtga cgctggtggc cctggtggat
     3061 gtggacaact atggccccat catcttggct gtgcatagga ctccagaggg cgtcatctac
     3121 ccttccatgt gctggatccg ggactccctg gtcagctaca tcaccaacct gggcctcttc
     3181 agcctggtgt ttctgttcaa catggccatg ctagccacca tggtggtgca gatcctgcgg
     3241 ctgcgccccc acacccaaaa gtggtcacat gtgctgacac tgctgggcct cagcctggtc
     3301 cttggcctgc cctgggcctt gatcttcttc tcctttgctt ctggcacctt ccagcttgtc
     3361 gtcctctacc ttttcagcat catcacctcc ttccaaggct tcctcatctt catctggtac
     3421 tggtccatgc ggctgcaggc ccggggtggc ccctcccctc tgaagagcaa ctcagacagc
     3481 gccaggctcc ccatcagctc gggcagcacc tcgtccagcc gcatctagaa gggcgaattc
     3541 tgcagatatc catcacactg gcggccgcag cacagtggtc gaaattctgg cgaaaggggg
     3601 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3661 aacgacggcc agtgaattgt aatacgactc actatagggc gaattaattc cggttatttt
     3721 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     3781 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     3841 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     3901 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     3961 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     4021 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     4081 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     4141 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4201 acgatgataa tatggccacc acccatatga aagaaaccgc tgctgctaaa atgaccgagt
     4261 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4321 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4381 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4441 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4501 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4561 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4621 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4681 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4741 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4801 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctgg tcgacaagca
     4861 tggggtaaag tactgttctc atcacatcat atcaaggtta tataccatca atattgccac
     4921 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct
     4981 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt
     5041 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat
     5101 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt
     5161 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat
     5221 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga
     5281 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt
     5341 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt
     5401 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt
     5461 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc
     5521 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct
     5581 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct
     5641 tttgggcttc tatatacatt ttagaatgag gttggcaagt tctcgacctg cagccaagct
     5701 tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     5761 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     5821 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     5881 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5941 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6001 aaggacctga aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     6061 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     6121 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     6181 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     6241 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     6301 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     6361 actccaatac tcctgaaata gttcattatg gacagcgcag aagagctggg gagaattaat
     6421 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6481 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6541 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6601 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6661 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6721 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7141 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7261 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7621 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7681 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7741 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7801 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7861 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7921 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7981 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8041 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8101 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8161 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8221 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8281 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8341 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8401 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8581 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8641 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     8701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     8761 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     8821 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     8881 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8941 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9001 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aagcagccca
     9061 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     9121 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     9181 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     9241 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
     9301 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     9361 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     9421 agaaaaaggg gggaa
//
