LOCUS       sequence-131215-        8618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4B 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 8618)
  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 8618)
  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: Sema4b-AP-His
FEATURES             Location/Qualifiers
     source          1..8618
                     /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)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2807..2818)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5198..5215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5241..5362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5278..5297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5332..5351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5400..5417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5571..6159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5651..5670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6330..7121,7122..7190))
                     /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     6953..6972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7191..7295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7363..7381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7419..7441)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7443..7463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7448..7778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7629..7764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7691..7710
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7915..7934
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8001..8456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8138..8159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8350..8369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8582..8604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8596..8613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8597..8613
                     /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 tgggccgggc
     1621 gagccgctcc gctgtcctgc ggcgcgcgct gctgctgctg ctgctactgc tgctgctgcg
     1681 gacgacgact acgcgggcgc tcggcccccg gatcagtgtg ccgctgggct ctgaagaaag
     1741 gctgattaga aaatttgaag ctgaaaacat ctccaactac acggcccttc tgctgagcca
     1801 ggatggcaag acgctgtatg tgggggcccg agaggccctc tttgcactta acagcaacct
     1861 cagcttcttg ccaggcgggg agtaccaaga gctgctgtgg agtgcagatg ctgacaggaa
     1921 gcagcagtgc agcttcaagg gcaaggaccc aaagcgtgac tgtcaaaact acatcaagat
     1981 cctcctgcca ctcaacagca gccacctgct cacctgtggc acggccgcct tcagccccct
     2041 gtgtgcttac attcacatag cgagctttac tttagcccaa gatgaggccg gcaatgtcat
     2101 tctggaggat ggcaagggtc gttgtccctt tgaccccaac ttcaagtcca cggctctggt
     2161 ggttgatggt gagctgtaca ctggaacagt cagtagcttc cagggaaacg acccagccat
     2221 ttcccggagc cagagttccc gccccaccaa gactgagagc tccctcaact ggctacaaga
     2281 ccctgccttt gtggcctcgg cctacgtccc cgagagcctg ggcagcccca taggtgatga
     2341 tgataagatc tacttcttct tcagcgagac gggccaggag tttgagttct ttgagaacac
     2401 catcgtgtcc cgagttgccc gagtctgtaa gggcgatgag ggtggagagc gggtgttgca
     2461 gcaacgctgg acctcctttc tcaaggctca gctcctgtgc tcccggcctg atgatggctt
     2521 tccctttaac gtgctacaag atgtcttcac cctgaacccc aaccctcagg attggcgcaa
     2581 gacccttttc tatggggtct ttacctccca gtggcacaga gggaccacag aaggctctgc
     2641 catctgcgtc ttcaccatga atgatgtgca gaaggccttt gacggcctgt acaagaaagt
     2701 aaacagagag acacagcagt ggtataccga gacccaccag gtgcccacac cgcggccggg
     2761 agcgtgcatt accaacagtg cccgggaacg gaagatcaac tcgtccctgc agctcccaga
     2821 ccgagtgctg aacttcctca aggatcactt cttgatggat gggcaggtcc gcagtcgcct
     2881 gctgctgctg cagcccagag cccgctacca gcgtgtggct gtgcaccgtg tgcctggcct
     2941 gcacagcact tatgatgtcc tatttctggg cactggtgat ggccgcctgc acaaagcagt
     3001 gaccctgagc tccagagtcc acatcattga ggagctgcag atcttccctc aaggacagcc
     3061 tgtgcagaac ctgctcttgg acagccatgg gggactgttg tatgcctcct cccattccgg
     3121 ggtggtgcaa gtgcccgtag ccaactgcag cctgtaccca acctgtggag actgcctcct
     3181 ggctcgagac ccctactgcg cctggactgg ctctgcctgc aggctcgcta gcctctacca
     3241 gcctgatctg gcctccaggc catggaccca ggacattgag ggtgccagtg tcaaggaact
     3301 ctgcaagaat tcctcataca aggcccggtt tcttgtgcca ggtaagccat gtaaacaagt
     3361 ccagatccaa ccaaacacag tgaacaccct ggcctgccca ctcctctcaa acctggccac
     3421 tcggctctgg gtgcacaatg gagccccagt caatgcctct gcctcctgcc gcgtgttacc
     3481 caccggggac ctgctgctgg tgggcagcca gcagggtttg ggggtgttcc agtgttggtc
     3541 gatagaagaa ggattccagc agcttgtggc cagctactgc ccagaggtga tggaggaggg
     3601 ggtaatggac caaaagaacc agcgtgatgg taccccagtc attatcaaca catcacgagt
     3661 gagtgcaccg gctggtggca gggccagctg gggtgcggac aagtcctacg gcgcgcctgg
     3721 atcttccgga atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     3781 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     3841 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     3901 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     3961 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     4021 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     4081 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     4141 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     4201 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     4261 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     4321 cgtgatccta ggtgggggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     4381 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     4441 gctggcgaag cgccagggtg cccggtacgt gtggaaccgc actgagctca tgcaggcttc
     4501 cctggacccg tctgtggccc atctcatggg tctctttgag cctggagaca tgaaatacga
     4561 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     4621 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     4681 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     4741 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     4801 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     4861 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     4921 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     4981 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     5041 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     5101 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     5161 gccccccgcc ggcaccaccg acgccgcgca cccgggtcat catcaccatc accattaatc
     5221 tagagatctt attaaagcag aacttgttta ttgcagctta taatggttac aaataaagca
     5281 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5341 ccaaactcat caatgtatct tatcatgtct ggtcgactct agctagactc ttccgcttcc
     5401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7321 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7381 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7441 gaggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta
     7501 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     7561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     7621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7681 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     7741 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     7801 ccattttcgt cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7861 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7921 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     7981 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     8041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     8101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     8161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     8221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     8281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     8341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     8401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     8461 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8521 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8581 tcccagtcac gacgttgtaa aacgacggcc agtgaatt
//
