LOCUS       sequence-60050-e        9224 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 9224)
  TITLE     BC-GPR56
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9224)
  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            "Unpublished"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-deltaSTP-GPR56
FEATURES             Location/Qualifiers
     source          1..9224
                     /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      3003..3012
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3445..3462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3446..3462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3469..3488
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3469..3487
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3538..3999
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3594..3611)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3821..3840
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4040..4636
                     /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(4040..4059)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4536..4556
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5537..6052
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6221..6237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6221..6237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6234..6256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6245..6261
                     /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(6269..6275,6276..6293,6294..6299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6314..6335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6452..6469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6623..7211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6703..6722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7382..8173,8174..8242))
                     /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     8005..8024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8243..8347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8415..8433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8471..8493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8593..8612
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9004..9023
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(9223..9224,1..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 cgccggaatt agatctaccg gtaagagtga ctccgtcgga
     1441 ggaaaatgac tccccagtcg ctgctgcaga cgacactgtt cctgctgagt ctgctcttcc
     1501 tggtccaagg tgcccacggc aggggccaca gggaagactt tcgcttctgc agccagcgga
     1561 accagacaca caggagcagc ctccactaca aacccacacc agacctgcgc atctccatcg
     1621 agaactccga agaggccctc acagtccatg cccctttccc tgcagcccac cctgcttccc
     1681 gatccttccc tgaccccagg ggcctctacc acttctgcct ctactggaac cgacatgctg
     1741 ggagattaca tcttctctat ggcaaggagc tcaaaaggga cctccagctg ctcagccagt
     1801 tcctgaagca tccccagaag gcctcaagga ggccctcggc tgcccccgcc agccagcagt
     1861 tgcagagcct ggagtcgaaa ctgacctctg tgagattcat gggggacatg gtgtccttcg
     1921 aggaggaccg gatcaacgcc acggtgtgga agctccagcc cacagccggc ctccaggacc
     1981 tgcacatcca ctcccggcag gaggaggagc agagcgagat catggagtac tcggtgctgc
     2041 tgcctcgaac actcttccag aggacgaaag gccggagcgg ggaggctgag aagagactcc
     2101 tcctggtgga cttcagcagc caagccctgt tccaggacaa gaattccagc caagtcctgg
     2161 gtgagaaggt cttggggatt gtggtacaga acaccaaagt agccaacctc acggagcccg
     2221 tggtgctcac cttccagcac cagctacagc cgaagaatgt gactctgcaa tgtgtgttct
     2281 gggttgaaga ccccacattg agcagcccgg ggcattggag cagtgctggg tgtgagaccg
     2341 tcaggagaga aacccaaaca tcctgcttct gcaaccactt gacctacttt gcagtgctga
     2401 tggtctcctc ggtggaggtg gacgccgtgc acaagcacta cctgagcctc ctctcctacg
     2461 tgggctgtgt cgtctctgcc ctggcctgcc ttgtcaccat tgccgcctac ctctgctcca
     2521 gggtgcccct gccgtgcagg aggaaacctc gggactacac catcaaggtg cacatgaacc
     2581 tgctgctggc cgtcttcctg ctggacacga gcttcctgct cagcgagccg gtggccctga
     2641 caggctctga ggctggctgc cgagccagtg ccatcttcct gcacttctcc ctgctcacct
     2701 gcctttcctg gatgggcctc gaggggtaca acctctaccg actcgtggtg gaggtctttg
     2761 gcacctatgt ccctggctac ctactcaagc tgagcgccat gggctggggc ttccccatct
     2821 ttctggtgac gctggtggcc ctggtggatg tggacaacta tggccccatc atcttggctg
     2881 tgcataggac tccagagggc gtcatctacc cttccatgtg ctggatccgg gactccctgg
     2941 tcagctacat caccaacctg ggcctcttca gcctggtgtt tctgttcaac atggccatgc
     3001 tagccaccat ggtggtgcag atcctgcggc tgcgccccca cacccaaaag tggtcacatg
     3061 tgctgacact gctgggcctc agcctggtcc ttggcctgcc ctgggccttg atcttcttct
     3121 cctttgcttc tggcaccttc cagcttgtcg tcctctacct tttcagcatc atcacctcct
     3181 tccaaggctt cctcatcttc atctggtact ggtccatgcg gctgcaggcc cggggtggcc
     3241 cctcccctct gaagagcaac tcagacagcg ccaggctccc catcagctcg ggcagcacct
     3301 cgtccagccg catctagaag ggcgaattct gcagatatcc atcacactgg cggccgcagc
     3361 acagtggtcg aaattctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     3421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     3481 ctatagggcg aattaattcc ggttattttc caccatattg ccgtcttttg gcaatgtgag
     3541 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3601 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3661 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3721 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3781 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3841 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     3901 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac
     3961 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacca cccatatgaa
     4021 agaaaccgct gctgctaaaa tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4081 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4141 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4201 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4261 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4321 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4381 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4441 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4501 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4561 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4621 ccgcaagccc ggtgcctggt cgacaagcat ggggtaaagt actgttctca tcacatcata
     4681 tcaaggttat ataccatcaa tattgccaca gatgttactt agccttttaa tatttctcta
     4741 atttagtgta tatgcaatga tagttctctg atttctgaga ttgagtttct catgtgtaat
     4801 gattatttag agtttctctt tcatctgttc aaatttttgt ctagttttat tttttactga
     4861 tttgtaagac ttctttttat aatctgcata ttacaattct ctttactggg gtgttgcaaa
     4921 tattttctgt cattctatgg cctgactttt cttaatggtt ttttaatttt aaaaataagt
     4981 cttaatattc atgcaatcta attaacaatc ttttctttgt ggttaggact ttgagtcata
     5041 agaaattttt ctctacactg aagtcatgat ggcatgcttc tatattattt tctaaaagat
     5101 ttaaagtttt gccttctcca tttagactta taattcactg gaattttttt gtgtgtatgg
     5161 tatgacatat gggttccctt ttatttttta catataaata tatttccctg tttttctaaa
     5221 aaagaaaaag atcatcattt tcccattgta aaatgccata tttttttcat aggtcactta
     5281 catatatcaa tgggtctgtt tctgagctct actctatttt atcagcctca ctgtctatcc
     5341 ccacacatct catgctttgc tctaaatctt gatatttagt ggaacattct ttcccatttt
     5401 gttctacaag aatatttttg ttattgtctt ttgggcttct atatacattt tagaatgagg
     5461 ttggcaagtt ctcgacctgc agccaagctt atcgataaaa taaaagattt tatttagtct
     5521 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     5581 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     5641 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5701 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     5761 agaaccatca gatgtttcca gggtgcccca aggacctgaa aatgaccctg tgccttattt
     5821 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     5881 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     5941 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     6001 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca
     6061 gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga
     6121 ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg
     6181 acagcgcaga agagctgggg agaattaatt cgtaatcatg gtcatagctg tttcctgtgt
     6241 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6301 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6361 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6481 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6541 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6601 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6661 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6721 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6781 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6841 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6901 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6961 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7021 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7081 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7141 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8341 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     8461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     8581 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     8641 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     8701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     8761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     8821 gtaaaacgac ggcgcaagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     8881 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     8941 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     9001 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     9061 cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt
     9121 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     9181 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaa
//
