LOCUS       sequence-309121-        6090 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses murine Vitronectin (VTN) protein with HA-tag at 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6090)
  AUTHORS   Bray ER, Yungher BJ, Levay K, Ribeiro M, Dvoryanchikov G, Ayupe AC, 
            Thakor K, Marks V, Randolph M, Danzi MC, Schmidt TM, Chaudhari N, 
            Lemmon VP, Hattar S, Park KK
  TITLE     Thrombospondin-1 Mediates Axon Regeneration in Retinal Ganglion 
            Cells.
  JOURNAL   Neuron. 2019 Aug 21;103(4):642-657.e7. doi: 
            10.1016/j.neuron.2019.05.044. Epub 2019 Jun 26.
  PUBMED    31255486
REFERENCE   2  (bases 1 to 6090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2019.05.044"; journalName: "Neuron"; date: 
            "2019-08-21- 21"; volume: "103"; issue: "4"; pages: "642-657.e7"
COMMENT     Sequence Label: pAAV.CMV.VTN.C-HA
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   49..189
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   60..189
                     /label=AAV2 ITR
     enhancer        265..568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        569..772
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     723..743
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     769..793
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          921..1299
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2848..2874
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2885..3361
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3084..3103)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3401..3541
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3401..3530
                     /label=AAV2 ITR
     rep_origin      3616..4071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3703..3722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3913..3934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4088..4107)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4207..4229
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4267..4285)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4353..4457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4458..4526,4527..5318)
                     /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     complement(4676..4695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5489..6077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5978..5997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
       61 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
      121 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      181 ggggttcctg cggccgcacg cgtctagtta ttaatagtaa tcaattacgg ggtcattagt
      241 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      301 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      361 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      421 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      481 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      541 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      601 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      661 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      721 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      781 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      841 ggaccgatcc agcctccgcg gattcgaatc ccggccggga acggtgcatt ggaacgcgga
      901 ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc acaaaaaatg
      961 ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc ctaatctctt
     1021 tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct aaagaataac
     1081 agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat ttctgcatat
     1141 aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc cagctaccat
     1201 tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag ctaggccctt
     1261 ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa cgtgctggtc
     1321 tgtgtgctgg cccatcactt tggcaaagaa ttgggattcg aacatcgatt gaattccccg
     1381 gggatcctct agagtcgacc tgcagaagct atggcacccc tgaggccctt tttcatacta
     1441 gccctggtgg catgggtttc tctggctgac caagagtcat gcaagggccg ctgcactcag
     1501 ggtttcatgg ccagcaagaa gtgtcagtgt gacgagcttt gcacttacta tcagagctgc
     1561 tgtgccgact acatggagca gtgcaagccc caagtaacgc ggggggacgt gttcactatg
     1621 ccagaggatg attattggag ctatgactac gtggaggagc ccaagaacaa taccaacacc
     1681 ggtgtgcaac ccgagaacac ctctccaccc ggtgacctaa atcctcggac ggacggcact
     1741 ctaaagccga cagccttcct agatcctgag gaacagccaa gcaccccagc gcctaaagtg
     1801 gagcaacagg aggagatcct aaggcccgac accactgatc aagggacccc tgagtttcca
     1861 gaggaagaac tgtgcagtgg aaagcccttt gacgccttca cggatctcaa gaatgggtcc
     1921 ctctttgcct tccgagggca gtactgctat gagctagatg agacggcagt gaggcctggg
     1981 taccccaaac ttatccaaga tgtctggggc attgagggcc ccatcgatgc tgccttcact
     2041 cgcatcaact gtcaggggaa gacctacttg ttcaagggta gtcagtactg gcgctttgag
     2101 gatggggtcc tggaccctgg ttatccccga aacatctccg aaggcttcag tggcatacca
     2161 gacaatgttg atgcagcgtt cgcccttcct gcccaccgtt acagtggccg ggaaagggtc
     2221 tacttcttca aggggaagca gtactgggag tacgaatttc agcagcaacc cagccaggag
     2281 gagtgcgaag gcagctctct gtcagccgtg tttgagcact ttgccttgct tcagcgggac
     2341 agctgggaga acattttcga actcctcttc tggggcagat cctctgatgg agccagagaa
     2401 ccccaattca tcagccggaa ctggcatggt gtgccaggga aagtggacgc tgctatggcc
     2461 ggccgcatct acgtcactgg ctccttatcc cactctgccc aagccaaaaa acagaagtct
     2521 aagcgtagaa gccgaaagcg ctatcgttca cgccgagggc gtggccacag acgcagccag
     2581 agctcgaact cccgtcgttc atcacgttca atctggttct ctttgttctc cagcgaggag
     2641 agtgggctag gaacctacaa caactatgat tatgatatgg actggcttgt acctgccacc
     2701 tgcgagccca ttcagagcgt ctatttcttc tctggagaca aatactaccg agtcaacctt
     2761 agaacccggc gagtggactc tgtgaatcct ccctacccac gctccattgc tcagtattgg
     2821 ctgggctgcc cgacctctga gaaagcttac ccatacgatg ttccagatta cgcttaggat
     2881 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     2941 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3001 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3061 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     3121 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     3181 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     3241 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     3301 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     3361 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3421 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3481 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3541 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3601 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3661 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3961 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4021 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4081 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4141 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4201 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4261 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4321 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4561 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4621 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4681 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4741 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4801 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4861 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4921 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4981 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5041 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5101 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5161 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5221 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5281 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5461 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5521 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5581 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5641 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5701 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5761 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5821 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5881 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5941 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6001 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6061 ggcggagcct atggaaaaac gccagcaacg
//
