LOCUS       sequence-60058-e        8313 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 8313)
  TITLE     BC-GPR56
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8313)
  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            "Unpublished"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-GPRC
FEATURES             Location/Qualifiers
     source          1..8313
                     /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"
     CDS             1423..1467
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     regulatory      2092..2101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2534..2551
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2535..2551
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2558..2577
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2558..2576
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2627..3088
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2683..2700)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2910..2929
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3129..3725
                     /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(3129..3148)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3625..3645
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4626..5141
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5310..5326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5310..5326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5323..5345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5334..5350
                     /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(5358..5364,5365..5382,5383..5388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5403..5424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5541..5558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5792..5811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6471..7262,7263..7331))
                     /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     7094..7113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7332..7436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7504..7522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7560..7582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7682..7701
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8093..8112
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8312..8313,1..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 gtatgtctgc acttctgatc
     1441 ctagctcttg ttggagctgc agttgctttg acctactttg cagtgctgat ggtctcctcg
     1501 gtggaggtgg acgccgtgca caagcactac ctgagcctcc tctcctacgt gggctgtgtc
     1561 gtctctgccc tggcctgcct tgtcaccatt gccgcctacc tctgctccag ggtgcccctg
     1621 ccgtgcagga ggaaacctcg ggactacacc atcaaggtgc acatgaacct gctgctggcc
     1681 gtcttcctgc tggacacgag cttcctgctc agcgagccgg tggccctgac aggctctgag
     1741 gctggctgcc gagccagtgc catcttcctg cacttctccc tgctcacctg cctttcctgg
     1801 atgggcctcg aggggtacaa cctctaccga ctcgtggtgg aggtctttgg cacctatgtc
     1861 cctggctacc tactcaagct gagcgccatg ggctggggct tccccatctt tctggtgacg
     1921 ctggtggccc tggtggatgt ggacaactat ggccccatca tcttggctgt gcataggact
     1981 ccagagggcg tcatctaccc ttccatgtgc tggatccggg actccctggt cagctacatc
     2041 accaacctgg gcctcttcag cctggtgttt ctgttcaaca tggccatgct agccaccatg
     2101 gtggtgcaga tcctgcggct gcgcccccac acccaaaagt ggtcacatgt gctgacactg
     2161 ctgggcctca gcctggtcct tggcctgccc tgggccttga tcttcttctc ctttgcttct
     2221 ggcaccttcc agcttgtcgt cctctacctt ttcagcatca tcacctcctt ccaaggcttc
     2281 ctcatcttca tctggtactg gtccatgcgg ctgcaggccc ggggtggccc ctcccctctg
     2341 aagagcaact cagacagcgc caggctcccc atcagctcgg gcagcacctc gtccagccgc
     2401 atctagaagg gcgaattctg cagatatcca tcacactggc ggccgcagca cagtggtcga
     2461 aattctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2521 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     2581 attaattccg gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2641 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2701 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2761 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2821 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2881 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2941 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3001 gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc acggggacgt
     3061 ggttttcctt tgaaaaacac gatgataata tggccaccac ccatatgaaa gaaaccgctg
     3121 ctgctaaaat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     3181 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     3241 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     3301 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     3361 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     3421 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     3481 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     3541 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     3601 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     3661 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     3721 gtgcctggtc gacaagcatg gggtaaagta ctgttctcat cacatcatat caaggttata
     3781 taccatcaat attgccacag atgttactta gccttttaat atttctctaa tttagtgtat
     3841 atgcaatgat agttctctga tttctgagat tgagtttctc atgtgtaatg attatttaga
     3901 gtttctcttt catctgttca aatttttgtc tagttttatt ttttactgat ttgtaagact
     3961 tctttttata atctgcatat tacaattctc tttactgggg tgttgcaaat attttctgtc
     4021 attctatggc ctgacttttc ttaatggttt tttaatttta aaaataagtc ttaatattca
     4081 tgcaatctaa ttaacaatct tttctttgtg gttaggactt tgagtcataa gaaatttttc
     4141 tctacactga agtcatgatg gcatgcttct atattatttt ctaaaagatt taaagttttg
     4201 ccttctccat ttagacttat aattcactgg aatttttttg tgtgtatggt atgacatatg
     4261 ggttcccttt tattttttac atataaatat atttccctgt ttttctaaaa aagaaaaaga
     4321 tcatcatttt cccattgtaa aatgccatat ttttttcata ggtcacttac atatatcaat
     4381 gggtctgttt ctgagctcta ctctatttta tcagcctcac tgtctatccc cacacatctc
     4441 atgctttgct ctaaatcttg atatttagtg gaacattctt tcccattttg ttctacaaga
     4501 atatttttgt tattgtcttt tgggcttcta tatacatttt agaatgaggt tggcaagttc
     4561 tcgacctgca gccaagctta tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     4621 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4681 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     4741 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4801 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     4861 atgtttccag ggtgccccaa ggacctgaaa atgaccctgt gccttatttg aactaaccaa
     4921 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4981 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta
     5041 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     5101 tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag
     5161 ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc
     5221 actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa
     5281 gagctgggga gaattaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7501 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     7561 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     7621 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     7681 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     7741 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     7801 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7861 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7921 gcgcaaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     7981 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     8041 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     8101 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     8161 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     8221 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     8281 agattttatt tagtctccag aaaaaggggg gaa
//
