LOCUS       Exported                8258 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses the Sema3F protein (truncated at cleavage site P1; ie, 
            short and contains no deletion in exon 3), C-terminally fused to 
            alkaline phosphatase + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    Sema3f(S, +)-AP-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  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 8258)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8258
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        105..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             4838..4855
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4881..5002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4918..4937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4972..4991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5040..5057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5211..5799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5291..5310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5970..6830)
                     /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     6593..6612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6831..6935)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7003..7021
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7059..7081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7083..7103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7088..7418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7269..7404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7331..7350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7555..7574
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7641..8096)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7778..7799)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7990..8009
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8222..8244
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8236..8253
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8237..8253
                     /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 tgcttgtcac
     1621 tgccttcatc ctctgggctt ccctgctgac cggggcctgg cctgctaccc ctatccagga
     1681 ccagcttcca gccacaccta gagtccgact gtcattcaaa gaacttaagg ccaccggtac
     1741 cgcccacttc ttcaactttc tgctcaacac tacagactac agaatcctgc tcaaggatga
     1801 ggaccatgac cgcatgtatg tgggcagcaa ggactacgtg ctgtccctgg acctgcatga
     1861 catcaaccga gagcccctta ttatccattg ggcagcctcc ccgcagcgca ttgaggagtg
     1921 catactgtca ggcaaggatg gcaatggaga gtgtggtaac ttcgtccggc tcatccagcc
     1981 ttggaaccga acacacctgt atgtgtgtgg gaccggtgcc tacaacccca tgtgcaccta
     2041 tgtgaaccgt ggccgtcgcg cacaggccct accgtggacc cagatgcagg tggtcagagg
     2101 ccgaggcagc agggccaccg atggtgccga ccgcccgacg cccacagccc cacgccagga
     2161 ttacatcttc tacctggagc ctgagaaact ggagtcagga aaaggcaaat gtccctatga
     2221 ccccaagctg gacacagcct cagccctcat caatgaggaa ctctatgcag gagtgtacat
     2281 cgattttatg ggtaccgatg cagccatttt ccgcacgctg gggaagcaga cagccatgcg
     2341 cacagatcag tacaactccc ggtggctcaa tgatccttcc ttcatacatg ctgagctcat
     2401 ccctgacagc gcagagcgca atgacgataa actctacttc ttcttccgag agcgctcggc
     2461 agaggcgcca cagaaccctg ctgtgtatgc ccgcatcggg cgcatctgcc ttaacgatga
     2521 cggtggccat tgctgcctgg tcaacaagtg gagcacattc ctgaaggctc ggctcgtctg
     2581 ctccgtgcca ggcgaggatg gtatcgagac ccacttcgac gaactccagg atgtgtttgt
     2641 ccagcagacc caggacatca ggaacccagt catttatgct gtctttacct cttcaggctc
     2701 tgtgttccga gggtctgcag tgtgtgtcta ctccatggct gatatccgca tggtcttcaa
     2761 tgggcctttt gctcacaagg agggccccaa ctatcaatgg atgcccttct caggaaagat
     2821 gccctatcct cggcctggca catgccctgg tggaaccttc acgccctcga tgaagtccac
     2881 taaggactac cctgatgaag tgatcaactt catgcgcact catccgctca tgtaccaggc
     2941 cgtctacccg ctccagcggc gacccctggt ggtccgtaca ggcgctccct accgcctcac
     3001 cactgtcgct gtggaccagg tggatgcagc tgatggacgc tatgaggtgc ttttcctggg
     3061 cacagaccgc gggacagtgc agaaggtcat cgtgctgccc aaggacgacc aggaggtgga
     3121 agagctgatg ctagaggaag tggaggtctt caaggaacca gcacctgtta agaccatgac
     3181 catctcttcc aagaggcaac aactgtatgt ggcctcagcc gtgggtgtca cacacctgag
     3241 cctgcaccgc tgccaggcct atggagctgc ctgtgccgac tgctgcctcg cccgcgatcc
     3301 ctactgtgcc tgggatggcc aggcctgctc tcgctacaca gcatcctccg gcgcgcctgg
     3361 atcttccgga atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     3421 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     3481 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     3541 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     3601 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     3661 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     3721 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     3781 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     3841 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     3901 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     3961 cgtgatccta ggtgggggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     4021 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     4081 gctggcgaag cgccagggtg cccggtacgt gtggaaccgc actgagctca tgcaggcttc
     4141 cctggacccg tctgtggccc atctcatggg tctctttgag cctggagaca tgaaatacga
     4201 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     4261 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     4321 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     4381 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     4441 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     4501 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     4561 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     4621 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     4681 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     4741 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     4801 gccccccgcc ggcaccaccg acgccgcgca cccgggtcat catcaccatc accattaatc
     4861 tagagatctt attaaagcag aacttgttta ttgcagctta taatggttac aaataaagca
     4921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4981 ccaaactcat caatgtatct tatcatgtct ggtcgactct agctagactc ttccgcttcc
     5041 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5101 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5401 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5641 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5881 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5941 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6001 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6061 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6121 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6181 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6241 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6301 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6361 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6421 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6481 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6541 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6601 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6661 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6721 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6781 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6841 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6961 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7021 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7081 gaggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta
     7141 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     7201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     7261 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7321 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     7381 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     7441 ccattttcgt cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7501 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7561 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     7621 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     7681 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7741 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7801 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7861 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7921 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7981 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     8041 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     8101 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8161 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8221 tcccagtcac gacgttgtaa aacgacggcc agtgaatt
//
