LOCUS       Exported                8546 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses the extracellular region of the Sema4G protein, 
            C-terminally fused to alkaline phosphatase + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    Sema4g-AP-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8546)
  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 8546)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8546
                     /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             5126..5143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5169..5290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5206..5225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5260..5279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5328..5345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5499..6087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5579..5598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6258..7118)
                     /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     6881..6900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7119..7223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7291..7309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7347..7369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7371..7391)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7376..7706
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7557..7692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7619..7638
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7843..7862
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7929..8384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8066..8087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8278..8297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8510..8532
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8524..8541
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8525..8541
                     /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 tgtgggggag
     1621 gctctggccc ctcctcttca gctttctcac agtaacagca gtgcctggac cctcactgcg
     1681 gagaccatcc cgagagctag atgccacccc acggctgacc atctcctatg aagagctctc
     1741 ccagattcgg cacttcaagg gccaaaccca gaactattca acactgctgc tggaggaggc
     1801 ttctgagagg ctgctggtgg gggctcgagg cgccctgttc tctctcagtg cccgtgacat
     1861 aagagatagg actcacaaag agatccactg ggaggcctcc ccagagatgc aaagcaagtg
     1921 tcatcagaaa gggaaaaaca accagaccga gtgtttcaac cacgtgcgat ttctacagcg
     1981 gctcaatgcc acccacttct atgcatgtgg gacccacgcc ttccagcccc tctgtgcagc
     2041 tattgatgct gaaaccttca tcttgcctac tagtttcgag gagggaaagg agaaatgtcc
     2101 ttatgacccg gcccggggct tcacaggcct tatcattgat ggaggcctct acacagccac
     2161 ccggtatgaa ttccggagca ttcctgatat ccgccggagc cgccacccac actctctgag
     2221 aactgaggag gcacccatgc actggctcaa tgatgctgag tttgtgttct ctgtgctggt
     2281 gcgggagagc aagacaagcg cagtgggtga tgatgacaag atttatttct tcttcatgga
     2341 gcgtgaggaa ggctccagca gcttcacaca gagtcgcagc agccaccgtg tggctagagt
     2401 agcccgtgtg tgcaagggag atctgggagg gaagaagatc ctacagaaga agtggacatc
     2461 cttcctgaag gctcgtctca tctgccacat cccacagtat gagacactac gtggggtctg
     2521 cagcctgaat gctgatacat catcccacac acatttctat gccgtcttca cactgacaac
     2581 acagtggaag acgctggagg cttcagccat ctgccgctat gacctggctg agattcaggc
     2641 tgtcttcaca ggccctttta tggagtacca ggatggtgcc cggcgctggg gccgctatga
     2701 gggtggggtg cctgaacccc ggcctggctc gtgcatcaca gattcattgc gcagccgagg
     2761 ctacaactca tcccaagact tgccatccct ggttctggac tttgtgaagt tgcatcccct
     2821 gatggctcgg cctgtggtgc ccacacgtgg gcggcccttg ctgctgaagc gcaatgtacg
     2881 ctacacacat ctcacaggaa cacacgtcag cacacctgct ggacccacct atgacctgct
     2941 ctttctgggc acagctgatg gctggatcca caaggccgta gtcctgggct ctgggatgca
     3001 cataattgaa gagatacaag tgttcaggga gccccagtct gtggacaatc tagttatctc
     3061 accaatgcag cacagcctct acgtgggggc tgcaagtgga gtcctccagt ttcctctgtc
     3121 cagctgctca cgctaccagt cctgctatga ctgcatcctg gcacgggacc cctactgtgg
     3181 ctgggactcc agcatccatg cctgcatggt ggccaccacc gtagccaaca ggacagaatt
     3241 gatacaggac atagagagag gaaatcgagg ctgtgagggc agcagggatg caggaccacc
     3301 accaccactc aagacccgat ctgtgctccg gggtgatgat gtcctcctgc cctgtgacca
     3361 gccgtctaac ctggcccggg ctttgtggct actcaatggg agcaagagcc tgagcgatgg
     3421 gcaggatggc taccgcgtgg gagtggatgg gctgctggta actgacacac aactggagca
     3481 cagtggcaac tatggctgct atgccgagga gaatggcctc cgtatgctgc tggcgtccta
     3541 cagcctcaca gtccggccag ccacccctgc tccagctcca caagcccctg ccacacctgg
     3601 ggcacagctg gcacatgaca tgaggatgtt ctatgtgggc gcgcctggat cttccggaat
     3661 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     3721 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     3781 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     3841 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     3901 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     3961 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     4021 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     4081 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     4141 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     4201 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     4261 tgggggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     4321 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     4381 ccagggtgcc cggtacgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     4441 tgtggcccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     4501 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     4561 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     4621 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     4681 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     4741 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     4801 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     4861 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     4921 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     4981 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     5041 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     5101 caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta gagatcttat
     5161 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5281 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     5341 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5401 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5461 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5521 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5581 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5641 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5701 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5761 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5881 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5941 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6001 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6061 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6121 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6181 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6241 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6301 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6361 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6421 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6481 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6541 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6601 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6661 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6721 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6781 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6841 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6901 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6961 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7021 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7081 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7141 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7201 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7261 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7321 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     7381 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     7441 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     7501 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     7561 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     7621 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     7681 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     7741 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7801 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     7861 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7921 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7981 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     8041 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     8101 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     8161 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     8221 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     8281 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     8341 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     8401 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8461 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8521 cgttgtaaaa cgacggccag tgaatt
//
