LOCUS       sequence-60060-e        8339 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 8339)
  TITLE     BC-GPR56
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8339)
  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            "Unpublished"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-Sig-GPRC
FEATURES             Location/Qualifiers
     source          1..8339
                     /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      2143..2152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2560..2577
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2561..2577
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2584..2603
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2584..2602
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2653..3114
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2709..2726)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2936..2955
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3155..3751
                     /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(3155..3174)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3651..3671
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4652..5167
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5336..5352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5336..5352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5349..5371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5360..5376
                     /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(5384..5390,5391..5408,5409..5414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6497..7288,7289..7357))
                     /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     7120..7139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7358..7462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7530..7548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7586..7608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7708..7727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8119..8138
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8338..8339,1..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 accggtcaag agtgactccg tcggaggaaa
     1441 atgactcccc agtcgctgct gcagacgaca ctgttcctgc tgagtctgct cttcctggtc
     1501 caaggtgccc acggcaggtt gacctacttt gcagtgctga tggtctcctc ggtggaggtg
     1561 gacgccgtgc acaagcacta cctgagcctc ctctcctacg tgggctgtgt cgtctctgcc
     1621 ctggcctgcc ttgtcaccat tgccgcctac ctctgctcca gggtgcccct gccgtgcagg
     1681 aggaaacctc gggactacac catcaaggtg cacatgaacc tgctgctggc cgtcttcctg
     1741 ctggacacga gcttcctgct cagcgagccg gtggccctga caggctctga ggctggctgc
     1801 cgagccagtg ccatcttcct gcacttctcc ctgctcacct gcctttcctg gatgggcctc
     1861 gaggggtaca acctctaccg actcgtggtg gaggtctttg gcacctatgt ccctggctac
     1921 ctactcaagc tgagcgccat gggctggggc ttccccatct ttctggtgac gctggtggcc
     1981 ctggtggatg tggacaacta tggccccatc atcttggctg tgcataggac tccagagggc
     2041 gtcatctacc cttccatgtg ctggatccgg gactccctgg tcagctacat caccaacctg
     2101 ggcctcttca gcctggtgtt tctgttcaac atggccatgc tagccaccat ggtggtgcag
     2161 atcctgcggc tgcgccccca cacccaaaag tggtcacatg tgctgacact gctgggcctc
     2221 agcctggtcc ttggcctgcc ctgggccttg atcttcttct cctttgcttc tggcaccttc
     2281 cagcttgtcg tcctctacct tttcagcatc atcacctcct tccaaggctt cctcatcttc
     2341 atctggtact ggtccatgcg gctgcaggcc cggggtggcc cctcccctct gaagagcaac
     2401 tcagacagcg ccaggctccc catcagctcg ggcagcacct cgtccagccg catctagtaa
     2461 tgagcggccg ccagcacagt ggtcgaaatt ctggcgaaag ggggatgtgc tgcaaggcga
     2521 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2581 ttgtaatacg actcactata gggcgaatta attccggtta ttttccacca tattgccgtc
     2641 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     2701 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     2761 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2821 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     2881 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     2941 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     3001 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt
     3061 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     3121 caccacccat atgaaagaaa ccgctgctgc taaaatgacc gagtacaagc ccacggtgcg
     3181 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3241 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3301 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3361 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3421 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3481 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3541 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     3601 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3661 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3721 cacctggtgc atgacccgca agcccggtgc ctggtcgaca agcatggggt aaagtactgt
     3781 tctcatcaca tcatatcaag gttatatacc atcaatattg ccacagatgt tacttagcct
     3841 tttaatattt ctctaattta gtgtatatgc aatgatagtt ctctgatttc tgagattgag
     3901 tttctcatgt gtaatgatta tttagagttt ctctttcatc tgttcaaatt tttgtctagt
     3961 tttatttttt actgatttgt aagacttctt tttataatct gcatattaca attctcttta
     4021 ctggggtgtt gcaaatattt tctgtcattc tatggcctga cttttcttaa tggtttttta
     4081 attttaaaaa taagtcttaa tattcatgca atctaattaa caatcttttc tttgtggtta
     4141 ggactttgag tcataagaaa tttttctcta cactgaagtc atgatggcat gcttctatat
     4201 tattttctaa aagatttaaa gttttgcctt ctccatttag acttataatt cactggaatt
     4261 tttttgtgtg tatggtatga catatgggtt cccttttatt ttttacatat aaatatattt
     4321 ccctgttttt ctaaaaaaga aaaagatcat cattttccca ttgtaaaatg ccatattttt
     4381 ttcataggtc acttacatat atcaatgggt ctgtttctga gctctactct attttatcag
     4441 cctcactgtc tatccccaca catctcatgc tttgctctaa atcttgatat ttagtggaac
     4501 attctttccc attttgttct acaagaatat ttttgttatt gtcttttggg cttctatata
     4561 cattttagaa tgaggttggc aagttctcga cctgcagcca agcttatcga taaaataaaa
     4621 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4681 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     4741 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     4801 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     4861 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaaatga
     4921 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4981 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     5041 gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     5101 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     5161 ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat
     5221 attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga
     5281 aatagttcat tatggacagc gcagaagagc tggggagaat taattcgtaa tcatggtcat
     5341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6781 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7081 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7501 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7561 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7621 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7681 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7741 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     7801 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7861 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7921 cccagtcacg acgttgtaaa acgacggcgc aaggaagcag cccagtagta ggttgaggcc
     7981 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     8041 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     8101 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     8161 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
     8221 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     8281 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaa
//
