LOCUS       sequence-131218-        8497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4F, isoform a protein,
            C-terminally fused to alkaline phosphatase + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8497)
  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 8497)
  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: Sema4f.a-AP-His
FEATURES             Location/Qualifiers
     source          1..8497
                     /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             5077..5094
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5120..5241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5157..5176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5211..5230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5279..5296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5450..6038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5530..5549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6209..7000,7001..7069))
                     /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     6832..6851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7070..7174)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7242..7260
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7298..7320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7322..7342)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7327..7657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7508..7643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7570..7589
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7794..7813
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7880..8335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8017..8038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8229..8248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8461..8483
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8475..8492
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8476..8492
                     /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 gccaccatgc tggccagggc cgagcggccc
     1621 cgcccgggcc cccggccgcc tccggtctct ctcttcccgc cgccgtcgtc gctgctgctg
     1681 ctgctgctgg cgatgctgag cgccccggtg tgcggccgcg tcccccgctc ggtgcccaga
     1741 acctcgctgc ccatctccga ggctgactcc tatctcaccc ggtttgcggc ccctcatacg
     1801 tacaattact ctgctctcct ggtggatcct gcttctcaca cactttacgt cggtgcacgg
     1861 gacagcatct tcgctttaac cctgcccttc tctggggaaa agcctcgaag gatcgactgg
     1921 atggtgcccg agactcacag acagaactgt aggaagaaag gcaagaaaga ggacgaatgt
     1981 cacaatttta tccagattct cgccattgcc aatgcctctc acctcctcac gtgtggcacc
     2041 ttcgcttttg atccgaagtg cggggttatt gatgtgtcca gtttccagca ggttgaaaga
     2101 cttgagagtg gccgggggaa atgtcctttt gagccagctc aacggtcagc agctgtaatg
     2161 gctgggggcg tcctctacac cgccactgtg aaaaacttcc tggggacaga gccgattatc
     2221 tcccgagctg tgggtcgagc tgaggactgg attcgaacag agaccttgtc atcctggctt
     2281 aatgccccag cctttgtcgc agctatggtc ctgagcccgg ctgagtgggg ggatgaagat
     2341 ggagacgatg aaatcttctt tttcttcacg gagacctccc gagtcttgga ctcatatgag
     2401 cgcatcaagg tcccacgagt ggcccgtgtg tgtgcggggg accttggggg ccggaagacc
     2461 cttcagcaga gatggacgac gtttctaaag gctgacctgc tgtgtccagg gcccgagcat
     2521 ggaagggcct cgggggttct gcaggatatg acagagcttc gacctcagcc tggcgcgggg
     2581 acccccctct tttatggcat cttttcctcc cagtgggaag gagccgccat ttctgctgtg
     2641 tgtgccttcc gaccccaaga catccgggca gtgctgaatg gtccctttag agagctaaaa
     2701 catgactgca acaggggact acctgtcatg gacaacgagg tgccccagcc cagacctgga
     2761 gagtgcatca ccaacaacat gaagttccag cagtttggat cctcactctc cctgccagac
     2821 cgtgtgctca cctttatcag agaccacccg ctcatggaca ggcccgtgtt tccagctgat
     2881 ggccgccccc tgctggtcac tacggataca gcctatctca gagttgtggc tcaccgggtg
     2941 accagcctct cagggaaaga atatgatgtg ctctacctag ggacagagga tgggcacctc
     3001 caccgggctg tgcgcatcgg agctcagctc agtgtcctgg aggatctggc cttgttcccg
     3061 gaaccacaac cggttgagag catgaaattg taccatgatt ggctcctggt gggctcccat
     3121 actgaggtga cacaagtgaa caccagcaac tgtggccgtc tccagagctg ctcagaatgt
     3181 atcctggccc aggatcccgt gtgtgcctgg agcttccggc tcgatgcttg tgtggcccat
     3241 gcaggcgagc accgtgggat ggttcaagac atagagtcag cagatgtctc ttctttgtgt
     3301 ccaaaagaac ctggagaaca tccggtagtg tttgaagttc ctgtggctac ggtgggccac
     3361 gtggtcctgc cgtgctcccc cagttctgcc tgggcatcct gtgtgtggca ccagcccagt
     3421 ggagtgactt cgctcacccc ccgaagggat ggactagagg tggtggtgac cccaggggcc
     3481 atgggggctt atgcttgcga gtgtcaggag ggtggagccg cccgtgtggt ggctgcttat
     3541 agcttggtgt ggggcagcca gagggggccc tcaaaccggg cccacaccgg cgcgcctgga
     3601 tcttccggaa tcatcccagt tgaggaggag aacccggact tctggaaccg cgaggcagcc
     3661 gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa gaacctcatc
     3721 atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat cctaaaaggg
     3781 cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt cccatatgtg
     3841 gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc cacagccacg
     3901 gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc agccgcccgc
     3961 tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag
     4021 aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc ctcgccagcc
     4081 ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg
     4141 gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat ggacattgac
     4201 gtgatcctag gtgggggccg aaagtacatg tttcgcatgg gaaccccaga ccctgagtac
     4261 ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt gcaggaatgg
     4321 ctggcgaagc gccagggtgc ccggtacgtg tggaaccgca ctgagctcat gcaggcttcc
     4381 ctggacccgt ctgtggccca tctcatgggt ctctttgagc ctggagacat gaaatacgag
     4441 atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc tgccctgcgc
     4501 ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg catcgaccat
     4561 ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc
     4621 attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt cactgccgac
     4681 cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat cttcgggctg
     4741 gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg aaacggtcca
     4801 ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag cgggagcccc
     4861 gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg cgaggacgtg
     4921 gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc
     4981 atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg cgacctggcg
     5041 ccccccgccg gcaccaccga cgccgcgcac ccgggtcatc atcaccatca ccattaatct
     5101 agagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca aataaagcaa
     5161 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5221 caaactcatc aatgtatctt atcatgtctg gtcgactcta gctagactct tccgcttcct
     5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5881 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7201 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7261 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7321 aggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat
     7381 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     7441 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     7501 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7561 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     7621 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     7681 cattttcgtc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7801 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7861 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     8041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     8101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     8161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     8221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     8281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     8341 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8401 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     8461 cccagtcacg acgttgtaaa acgacggcca gtgaatt
//
