LOCUS       Exported                7754 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses the extracellular region of the Sema4G protein, 
            C-terminally fused to the Fc region of human IgG1 + 6X histidine 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    Sema4g-Fc-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7754)
  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 7754)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7754
                     /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             4334..4351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4377..4498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4414..4433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4468..4487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4536..4553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4707..5295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4787..4806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5466..6326)
                     /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     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6499..6517
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6555..6577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6579..6599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6584..6914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6765..6900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6827..6846
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7051..7070
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7137..7592)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7274..7295)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7486..7505
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7718..7740
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7732..7749
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7733..7749
                     /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 gcgcctgaca aaactcacac
     3661 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     3721 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     3781 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     3841 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     3901 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     3961 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     4021 accacaggtg tataccctgc ccccatcccg ggatgagctg accaagaacc aggtcagcct
     4081 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     4141 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     4201 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     4261 ctccgtgatg catgaggctc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     4321 gggtaaaggc ggccatcatc accatcacca ttaatctaga gatcttatta aagcagaact
     4381 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4441 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4501 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg
     4561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6481 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     6541 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg
     6601 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     6661 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6721 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6841 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6901 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca
     6961 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7021 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7081 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     7141 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7201 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7261 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7321 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7381 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7441 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7501 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7561 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     7621 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7681 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     7741 acggccagtg aatt
//
