LOCUS       sequence-131319-        6986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the RGMa protein, C-terminally
            fused to the Fc region of human IgG1 + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 6986)
  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: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Rgma-Fc-His
FEATURES             Location/Qualifiers
     source          1..6986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             3566..3583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3609..3730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3646..3665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3700..3719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3768..3785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3939..4527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4019..4038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4698..5489,5490..5558))
                     /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     5321..5340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5559..5663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5731..5749
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5787..5809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5811..5831)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5816..6146
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5997..6132
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6059..6078
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6283..6302
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6369..6824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6506..6527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6718..6737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6950..6972
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6964..6981
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6965..6981
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tgcagccgcc
     1621 aagggagagg ctagtggtaa caggccgagc tggatggatg ggtatgggga gaggggcagg
     1681 acgttcagcc ctgggattgt ggccgaccct cgccttcctt ctctgcagct tccccgcagc
     1741 catctccccc tgcaaaatcc tcaagtgcaa ctctgagttc tggagcgcca cgtcatcagg
     1801 cagccacgcc cctgcctccg acgacgtgcc ggagttctgt gcagccctgc gcacctacgc
     1861 cctgtgcact cggcggacag cccgcacctg ccggggcgac ctggcttacc actcggctgt
     1921 ccatggcata gaggacctta tgagccagca caactgctcc aaggacggcc ccacctcaca
     1981 gccgcgagtg cgcacgctcc cgccagctgg ggacagccag gagcgctcgg atagccccga
     2041 gatctgccac tatgagaaga gtttccacaa gcactcagct gcccccaact acactcactg
     2101 cggcctcttt ggggacccac acctcaggac tttcacagac cacttccaga catgcaaggt
     2161 gcaaggcgcc tggcctctca tcgacaataa ttacctgaac gtgcaagtca ccaatacacc
     2221 tgtgctgccg ggctccgcag ctaccgccac cagcaagctc accatcatct tcaagaactt
     2281 ccaagagtgt gtggaccaga aagtctacca agctgaaatg gacgaacttc cgtctgcctt
     2341 tgcggacggc tccaaaaatg ggggagataa acacggggcc aacagcctga agatcacaga
     2401 gaaggtgtcg ggccagcacg tggagatcca ggccaagtac atcggcacca ccatcgtggt
     2461 gcgacaggtg ggccgctacc tgacctttgc cgtccggatg cccgaggagg tagtcaacgc
     2521 cgtggaggac cgtgacagcc aaggcctcta cctctgcctg cggggctgcc cgctcaacca
     2581 gcagatcgac ttccaggctt tccgtgccaa cgctgaaagc cctcgcaggc cagcagccgc
     2641 cagtccctct cccgtggtcc ccgagacatt cccctatgag acagccgtgg ccaagtgcaa
     2701 agagaagctg cccgtagaag acctgtacta ccaggcctgt gtcttcgacc tcctcacgac
     2761 tggtgatgtg aacttcacgc tggctgccta ctatgctttg gaggatggca agatgctcca
     2821 ctccaacaag gacaagctgc atctgtttga aaggactcgg gagctgccag gaactagtga
     2881 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     2941 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     3001 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     3061 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     3121 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     3181 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     3241 gcagccccga gaaccacagg tgtataccct gcccccatcc cgggatgagc tgaccaagaa
     3301 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     3361 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     3421 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     3481 cgtcttctca tgctccgtga tgcatgaggc tctgcacaac cactacacgc agaagagcct
     3541 ctccctgtct ccgggtaaag gcggccatca tcaccatcac cattaatcta gagatcttat
     3601 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3661 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3721 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     3781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5701 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     5821 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     5881 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5941 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6001 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6061 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     6121 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     6181 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6241 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6301 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6361 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6481 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6541 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6601 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6661 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6721 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     6781 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     6841 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     6901 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6961 cgttgtaaaa cgacggccag tgaatt
//
