LOCUS       Exported                6224 bp ds-DNA     circular SYN 28-APR-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 28, 2022 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6224
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          664..665
                     /organism="synthetic DNA construct"
                     /plasmid="NEBuilder_Assembly"
                     /note="color: #ffffff"
     source          664..665
                     /organism="synthetic DNA construct
                     /mol_type=""other DNA
                     /note=""pCAG"
                     /note="color: #ffffff"
     source          672..2320
                     /organism="synthetic DNA construct"
                     /plasmid="NEBuilder_Assembly"
                     /note="color: #ffffff"
     source          672..1401
                     /organism="synthetic DNA construct
                     /mol_type=""other DNA
                     /note=""pCAG"
                     /note="color: #ffffff"
     source          1402..2026
                     /organism="synthetic DNA construct
                     /mol_type=""other DNA
                     /note=""IRES"
                     /note="color: #ffffff"
     source          2027..2320
                     /organism="synthetic DNA construct
                     /mol_type=""other DNA
                     /note=""mSnca"
                     /note="color: #ffffff"
     misc_feature    1..141
                     /label=L-ITR
                     /note="color: #a6acb3"
     primer_bind     complement(23..34)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     84..95
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     150..173
                     /label=MluI-Syn-F
                     /note="color: black; sequence: ATTACGCGTtaactgcagagggccctg"
     promoter        156..626
                     /label=Syn
                     /note="color: #ffffff"
     primer_bind     509..527
                     /label=Syn-Seq
                     /note="color: black; sequence: acgcgaggcgcgagatagg"
     regulatory      673..682
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             679..1398
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  679 ..  681 / #05fd14
                        2:  682 ..  684 / #05fd14 / 1a
                        3:  685 .. 1398 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1376..1404)
                     /label=GFP-rev
                     /note="color: black; sequence: 
                     ctggctttacttgtacagctcgtccatgc"
     primer_bind     1385..1433
                     /label=IRES_fwd
                     /note="3'Tm=76.1 3'Ta(annealing temp)=72.0"
                     /note="color: black; sequence: 
                     agctgtacaagtaaagcCAGTGTGGTGGTACGTAGGAATTCTCCCTCCC"
     misc_feature    1426..1992
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #a6acb3"
     primer_bind     complement(1515..1534)
                     /label=IRES-R
                     /note="color: black; sequence: cctcacattgccaaaagacg"
     primer_bind     complement(1997..2030)
                     /label=IRES_rev
                     /note="3'Tm=80.3 3'Ta(annealing temp)=72.0"
                     /note="color: black; sequence: 
                     tggcgtcgactctagaGGATCCCGGGCCCGCGGT"
     primer_bind     2007..2044
                     /label=mSnca_fwd
                     /note="3'Tm=63.8 3'Ta(annealing temp)=61.7"
                     /note="color: black; sequence: 
                     cgggatcctctagagtcgacGCCACCATGGATGTGTTC"
     regulatory      2027..2036
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             2033..2320
                     /codon_start=1
                     /label=mSnca(1-95)
                     /note="This feature has 2 segments:
                        1: 2033 .. 2317 / #cc99ff
                        2: 2318 .. 2320 / #cc99ff"
                     /translation="MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTK
                     EGVVHGVTTVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGNIAAATGFV"
     misc_feature    2341..2949
                     /label=WPRE
                     /note="color: #a6acb3"
     primer_bind     complement(2404..2424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     primer_bind     complement(2564..2583)
                     /label=seq-wpre-R (Sui)
                     /note="sw primer box2 #1392"
                     /note="color: black; sequence: gttgcgtcagcaaacacagt"
     misc_feature    2953..2955
                     /label=AfeI
                     /note="color: #ff99cc"
     polyA_signal    2969..3447
                     /label=hGH polyA
                     /note="color: #a6acb3"
     primer_bind     complement(2984..3001)
                     /label=bGH-R
                     /note="color: black; sequence: gcactggggaggggtcac"
     misc_feature    3487..3627
                     /label=R-ITR
                     /note="color: #a6acb3"
     primer_bind     complement(3533..3544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     3594..3605
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      3719..4025
                     /label=f1 Origin
                     /note="color: #ffff00"
     primer_bind     complement(3789..3808)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     3999..4020
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     promoter        4439..4543
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4544..5404
                     /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: 4544 .. 4612 / #ccffcc / signal sequence
                        2: 4613 .. 5404 / #ccffcc
                     Cleavage site after base 4612"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    4544..5401
                     /label=AmpR
                     /note="color: #a6acb3"
     primer_bind     complement(4762..4781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      5552..6219
                     /label=pUC ori
                     /note="color: #ffff00"
     primer_bind     6064..6083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttaact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcgtccg gagctattag caatatgaaa cctcttaggc
      661 gcgccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1261 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1381 gacgagctgt acaagtaaag ccagtgtggt ggtacgtagg aattctccct cccccccccc
     1441 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1501 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     1561 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     1621 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     1681 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     1741 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     1801 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     1861 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta cgtgtgttta
     1921 gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     1981 acacgatgat aatggtaccg cgggcccggg atcctctaga gtcgacgcca ccatggatgt
     2041 gttcatgaaa ggactttcaa aggccaagga gggagttgtg gctgctgctg agaaaaccaa
     2101 gcagggtgtg gcagaggcag ctggaaagac aaaagaggga gtcctctatg taggttccaa
     2161 aactaaggaa ggagtggttc atggagtgac aacagtggct gagaagacca aagagcaagt
     2221 gacaaatgtt ggaggagcag tggtgactgg tgtgacagca gtcgctcaga agacagtgga
     2281 gggagctggg aatatagctg ctgccactgg ctttgtctaa gctagcgaat tcgatatcaa
     2341 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2401 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2461 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2521 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2581 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2641 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2701 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2761 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2821 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2881 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2941 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3001 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3061 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3121 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3181 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3241 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3301 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3361 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3421 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3481 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3541 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3601 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3661 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3901 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4021 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4141 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4201 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4261 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4321 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
