LOCUS       sequence-135786-        6929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the JAM-4, isoform 3 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 6929)
  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 6929)
  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: Jam4.3-AP-His
FEATURES             Location/Qualifiers
     source          1..6929
                     /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             3509..3526
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3552..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3589..3608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3643..3662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3711..3728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3882..4470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3962..3981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4641..5432,5433..5501))
                     /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     5264..5283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5502..5606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5674..5692
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5730..5752)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5754..5774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5759..6089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5940..6075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6002..6021
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6226..6245
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6312..6767)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6449..6470)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6661..6680
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6893..6915
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6907..6924
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6908..6924
                     /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 tggagggcag
     1621 ctggagagat gtcctggctg tgctggtcat cctggctcag ctgacagctt ccggatccag
     1681 ttatcagatc atagaaggtc ctcagaatgt aacagtccta aaggactcag aggctcactt
     1741 caactgcacc gtgactcacg gctggaagct tctcatgtgg actcttaacc aaatggtggt
     1801 gctgagtctc accacccaag gacccatcat caccaacaac cgcttcacct atgccagtta
     1861 caacagcact gacagcttca tctcggagtt gatcatccat gatgtgcagc ccagtgactc
     1921 gggatccgtg caatgcagcc tgcagaacag ccatgggttt ggatctgcct tcctctcagt
     1981 gcaagacagt attggagagg aaggcccagc actgccgacc ggcgcgcctg gatcttccgg
     2041 aatcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     2101 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     2161 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     2221 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     2281 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     2341 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     2401 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     2461 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     2521 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     2581 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     2641 aggtgggggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     2701 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     2761 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     2821 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     2881 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     2941 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     3001 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     3061 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     3121 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     3181 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     3241 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     3301 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     3361 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     3421 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     3481 cggcaccacc gacgccgcgc acccgggtca tcatcaccat caccattaat ctagagatct
     3541 tattaaagca gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3661 tcaatgtatc ttatcatgtc tggtcgactc tagctagact cttccgcttc ctcgctcact
     3721 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3781 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3841 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3901 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3961 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4021 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4081 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4141 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4201 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4261 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4321 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4381 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4441 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4501 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4561 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4621 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4681 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4741 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4801 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4861 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4921 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4981 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5041 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5101 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5161 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5221 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5281 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5341 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5401 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5461 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5521 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5581 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5641 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5701 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggaggaatgt
     5761 gtgtcagtta gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca
     5821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6001 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     6061 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     6121 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6181 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6241 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6301 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6361 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6421 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6481 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     6541 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     6601 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     6661 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     6721 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     6781 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6841 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6901 cgacgttgta aaacgacggc cagtgaatt
//
