LOCUS       Exported                5118 bp ds-DNA     circular UNA 04-DEC-2025
DEFINITION  natural linear DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5118)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 4, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Template 155A digested with BamHI, NheI 
            f1:gaagcttGCCACCatgCACCTGAGAATCCACGCGAGACGG 
            f2:gtcgtgcctgagagcgcagtcgagaagaagcttGCCACCatgCACCTGAGAATC 
            r1:GTATGGCTCCGGCcAGCTTGCGCCGCGC
            f2:GCGCGGCGCAAGCTgGCCGGAGCCATAC
            r2:CTTATCGTCGTCATCCTTGTAGTCggatc
FEATURES             Location/Qualifiers
     source          1..5118
                     /organism="unspecified"
                     /mol_type="genomic DNA"
                     /note="color: #ffffff"
     misc_feature    1..130
                     /label=ITR
                     /note="color: #a6acb3"
     misc_feature    151..628
                     /label=synapsin promoter
                     /note="color: #a6acb3"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    637..642
                     /label=HindIII
                     /note="color: #a6acb3"
     misc_feature    649..651
                     /label=Ig k-chain signal sequence
                     /note="color: #a6acb3"
     misc_feature    join(652..810,838..975)
                     /label=NRX3b
                     /label=NRX3b(1)
                     /note="This feature has 2 segments:
                        1: 652 .. 810 / #a6acb3
                        2: 838 .. 975 / #a6acb3"
     misc_feature    811..837
                     /label=HA tag
                     /note="color: #a6acb3"
     CDS             811..837
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     misc_feature    976..984
                     /label=NRX3b
                     /note="color: #a6acb3"
     misc_feature    985..996
                     /label=NRX3b
                     /label=NRX3b(2)
                     /note="color: #a6acb3"
     misc_feature    997
                     /label=10 aa linker
                     /label=10 aa linker(2)
                     /note="color: #a6acb3"
     misc_feature    998..1014
                     /label=10 aa linker
                     /label=10 aa linker(1)
                     /note="color: #a6acb3"
     misc_feature    998..1014
                     /label=annealing region for overlap
                     /note="color: #a6acb3"
     misc_feature    1015..1023
                     /label=10 aa linker
                     /note="color: #a6acb3"
     misc_feature    1063..1104
                     /label=V5 tag
                     /note="color: #a6acb3"
     CDS             1063..1104
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1105..1110
                     /label=NheI
                     /note="color: #a6acb3"
     CDS             1111..2067
                     /label=BirA (humanized)
                     /note="color: #993366; direction: RIGHT"
     misc_feature    1300..1302
                     /label=Q65P
                     /note="color: #a6acb3"
     misc_feature    1366..1368
                     /label=I87V
                     /note="color: #a6acb3"
     misc_feature    1450..1452
                     /label=G115G
                     /note="color: #a6acb3"
     misc_feature    1459..1461
                     /label=R118S
                     /note="color: #a6acb3"
     misc_feature    1525..1527
                     /label=E141K
                     /note="color: #a6acb3"
     misc_difference 1528..1530
                     /label=Q142R
                     /note="color: #a6acb3"
     misc_feature    1552..1554
                     /label=S150G
                     /note="color: #a6acb3"
     misc_feature    1555..1557
                     /label=L151P
                     /note="color: #a6acb3"
     misc_feature    1582..1584
                     /label=V160A
                     /note="color: #a6acb3"
     misc_feature    1678..1680
                     /label=T192A
                     /note="color: #a6acb3"
     misc_difference 1684..1686
                     /label=K194I
                     /note="color: #a6acb3"
     misc_feature    1729..1731
                     /label=M209V
                     /note="color: #a6acb3"
     misc_feature    1825..1827
                     /label=M241T
                     /note="color: #a6acb3"
     misc_feature    1891..1893
                     /label=S263P
                     /note="color: #a6acb3"
     misc_structure  1975..1977
                     /label=G291G
                     /note="color: #a6acb3"
     misc_feature    2017..2019
                     /label=I305V
                     /note="color: #a6acb3"
     misc_feature    2068..2078
                     /label=NES
                     /label=NES(1)
                     /note="color: #a6acb3"
     CDS             2074..2100
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /note="color: #993366"
                     /translation="LPPLERLTL"
     misc_feature    2079
                     /label=NES
                     /label=NES(2)
                     /note="color: #a6acb3"
     misc_feature    2080..2103
                     /label=NES
                     /note="color: #a6acb3"
     misc_feature    complement(2104..2106)
                     /label=double stop
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2110..2115
                     /label=EcoRI
                     /note="color: #a6acb3"
     misc_feature    2122..2353
                     /label=PolyA
                     /note="color: #a6acb3"
     polyA_signal    2146..2353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    2382..2522
                     /label=ITR
                     /label=ITR(1)
                     /note="color: #a6acb3"
     repeat_region   2382..2522
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      2596..3051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3333..3437
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3438..4298
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3438 .. 3506 / #ccffcc / signal sequence
                        2: 3507 .. 4298 / #ccffcc
                     Cleavage site after base 3506"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4469..5057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaagaagc ttgccaccat gcacctgaga
      661 atccacgcga gacggagccc tcctcgccgg ccggcctgga cgcttgggat ctggttcctg
      721 ttctggggat gtatcgtcag ctctgtatgg agtcaattgt cttctaatgt agcttcctcc
      781 tcctccacct cttcctcgcc ggggtctcac tacccatacg atgtgccaga ttacgccaac
      841 cccacggagc cgggaatcag acgggttccg ggggcctcag aggtgatccg ggagtcgagc
      901 agcacaacag ggatggtcgt cggcattgtg gctgctgccg ccctctgcat cttgatcctc
      961 ctgtacgcca tgtacaagta caggaacagg gacgagggct cgggctcgac ctcgggctcg
     1021 ggcagcggtg gctctggagg ttctggtggc agctctggag gtggcaagcc catccccaac
     1081 cccctgctgg gcctggacag caccgctagc aaagacaata ctgtgcctct gaagctgatc
     1141 gctctcctgg ctaatggcga gttccatagt ggcgaacagc tgggagaaac cctgggcatg
     1201 tccagggccg ctatcaacaa gcacattcag actctgcgcg actggggcgt ggacgtgttc
     1261 accgtgcccg gaaagggcta ctctctgccc gagcctatcc cgctgctgaa cgctaaacag
     1321 attctgggac agctggacgg cgggagcgtg gcagtcctgc ctgtggtcga ctccaccaat
     1381 cagtacctgc tggatcgaat cggcgagctg aagagtgggg atgcttgcat tgcagaatat
     1441 cagcaggcag ggagaggaag cagagggagg aaatggttct ctccttttgg agctaacctg
     1501 tacctgagta tgttttggcg cctgaagcgg ggaccagcag caatcggcct gggcccggtc
     1561 atcggaattg tcatggcaga agcgctgcga aagctgggag cagacaaggt gcgagtcaaa
     1621 tggcccaatg acctgtatct gcaggataga aagctggcag gcatcctggt ggagctggcc
     1681 ggaataacag gcgatgctgc acagatcgtc attggcgccg ggattaacgt ggctatgagg
     1741 cgcgtggagg aaagcgtggt caatcagggc tggatcacac tgcaggaagc agggattaac
     1801 ctggacagga atactctggc cgctacgctg atccgagagc tgcgggcagc cctggaactg
     1861 ttcgagcagg aaggcctggc tccatatctg ccacggtggg agaagctgga taacttcatc
     1921 aatagacccg tgaagctgat cattggggac aaagagattt tcgggattag ccgggggatt
     1981 gataaacagg gagccctgct gctggaacag gacggagtta tcaaaccctg gatgggcgga
     2041 gaaatcagtc tgcggtctgc cgaaaagctg cagctgcctc ccctggagcg cctgaccctg
     2101 gactaatagg aattcctcga gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     2161 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2221 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2281 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2341 ggcatgctgg ggaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2401 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2461 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2521 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtattcaca ccgcatacgt
     2581 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     2941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3001 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3061 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3121 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3181 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3241 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3301 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3361 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3421 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3481 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3541 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3601 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3661 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3721 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3781 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3841 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3901 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3961 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4021 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4081 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4141 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4201 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4261 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4321 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4381 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4441 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4501 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4561 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4621 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4681 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4741 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4801 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4861 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4921 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4981 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5041 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5101 ggccttttgc tcacatgt
//
