LOCUS       sequence-131217-        8714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4D 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 8714)
  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 8714)
  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: Sema4d-AP-His
FEATURES             Location/Qualifiers
     source          1..8714
                     /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             5294..5311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5337..5458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5374..5393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5428..5447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5496..5513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6426..7217,7218..7286))
                     /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     7049..7068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7287..7391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7459..7477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7515..7537)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7539..7559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7544..7874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7725..7860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7787..7806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8011..8030
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8097..8552)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8234..8255)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8446..8465
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8678..8700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8692..8709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8693..8709
                     /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 tgaggatgtg
     1621 tgcccccgtt agggggctgt tcttggccct ggtggtagtg ttgagaaccg cggtggcatt
     1681 tgcacctgtg cctcggctca cctgggaaca tggagaggta ggtctggtgc agtttcacaa
     1741 gccaggcatc tttaactact cggccttgct gatgagtgag gacaaagaca ctctgtatgt
     1801 aggcgcccgg gaagcagtct ttgcagtgaa tgcgctgaac atctctgaga agcaacatga
     1861 ggtatattgg aaggtctctg aagacaaaaa atccaagtgt gcagagaagg ggaaatcaaa
     1921 gcagacggaa tgcctaaact acattcgagt actacagcca ctaagcagca cttccctcta
     1981 tgtgtgtggg accaatgcgt tccagcccac ctgtgaccac ctgaacttga catccttcaa
     2041 gtttctgggg aaaagtgaag atggcaaagg aagatgcccc ttcgaccccg cccacagcta
     2101 cacatcagtc atggttgggg gcgagctcta ctctgggacg tcctataatt tcttgggcag
     2161 tgaacccatc atctctcgaa actcttccca cagtcccttg aggacggagt atgccatccc
     2221 gtggctgaac gagcctagct tcgtctttgc tgacgtgatc cagaaaagcc cagatggtcc
     2281 ggagggtgaa gatgacaagg tctacttctt ttttacggag gtatccgtgg agtacgaatt
     2341 cgtcttcaag ttgatgatcc cgcgagttgc cagggtgtgc aagggcgacc agggcggcct
     2401 gcggactttg caaaaaaagt ggacctcctt cctaaaggcc aggctgatct gctccaagcc
     2461 agacagtggc ctggtcttca acatacttca ggatgtgttt gtgctgaggg ccccgggcct
     2521 caaggagcct gtgttctatg cggtcttcac cccacagctg aacaatgtgg gtctgtcagc
     2581 ggtgtgcgcc tacacactgg ccacggtgga ggcagtcttc tcccgtggaa agtacatgca
     2641 gagtgccaca gtggagcagt ctcacaccaa gtgggtgcgc tacaatggcc cagtgcccac
     2701 tccccgacct ggagcgtgta tcgacagtga ggcccgggca gccaactaca ccagctcctt
     2761 gaatctccca gacaaaacac tgcagtttgt aaaagaccac cctttgatgg atgactcagt
     2821 gaccccgata gacaacagac ccaagctgat caaaaaagat gtaaactaca cccagatagt
     2881 ggtagacagg acccaggccc tggatgggac tttctacgac gtcatgttca tcagcacaga
     2941 ccggggagct ctgcataaag cagtcatcct cacaaaagag gtgcatgtca tcgaggagac
     3001 ccaactcttc cgggactctg aaccggtcct aactctgctg ctatcgtcaa agaaggggag
     3061 gaagtttgtc tatgcaggct ccaactctgg agtggtccaa gcgcccctgg cattctgcga
     3121 aaagcacggt agctgtgaag actgtgtgtt agcacgggac ccctactgtg cctggagccc
     3181 agccatcaag gcctgtgtta ccctgcacca ggaagaggcc tccagcaggg gctggattca
     3241 ggacatgagc ggtgacacat cctcatgcct ggataagagt aaagaaagtt tcaaccagca
     3301 ttttttcaag cacggcggca cagcggaact caaatgtttc caaaagtcca acctagcccg
     3361 ggtggtatgg aagttccaga atggcgagtt gaaggccgca agtcccaagt acggctttgt
     3421 gggcaggaag cacctgctca tcttcaacct gtcggacgga gacagcggcg tgtaccagtg
     3481 cctgtcagag gaaagggtga ggaataaaac ggtctcccag ctgctggcca agcacgttct
     3541 ggaagtgaag atggtacctc ggaccccccc ctcacctacc tcagaggatg ctcagacaga
     3601 aggtagtaag atcacatcca aaatgccggt tgcatctacc caggggtcct ctccccctac
     3661 cccggctctg tgggcaacct cccccagagc cgccacccta cctcccaagt cctcctccgg
     3721 cacatcctgt gaaccaaaga tggtcatcaa cacggtcccc cagctccact cagagaagac
     3781 ggtgtatctc aagtccagtg acaacggcgc gcctggatct tccggaatca tcccagttga
     3841 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     3901 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     3961 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     4021 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     4081 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     4141 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     4201 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     4261 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     4321 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     4381 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg ggggccgaaa
     4441 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     4501 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     4561 gtacgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tggcccatct
     4621 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     4681 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     4741 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     4801 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     4861 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     4921 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     4981 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     5041 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     5101 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     5161 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     5221 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     5281 cgcgcacccg ggtcatcatc accatcacca ttaatctaga gatcttatta aagcagaact
     5341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5461 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     7441 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     7501 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg
     7561 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     7621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     7681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7801 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7861 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca
     7921 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7981 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     8041 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     8101 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     8161 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     8221 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     8281 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     8341 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     8401 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8461 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8521 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     8581 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8641 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8701 acggccagtg aatt
//
