LOCUS       Exported                7842 bp ds-DNA     circular SYN 31-JUL-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7842)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Feb 26, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7842
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        232..819
                     /label=CMV promoter
                     /note="color: #ffffff"
     primer_bind     769..789
                     /label=CMV forward primer
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    804..807
                     /label=TATA box
                     /note="color: #00ffff"
     misc_feature    830
                     /label=putative transcriptional start
                     /note="color: #ff0000"
     primer_bind     863..882
                     /label=T7 primer
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        863..879
                     /label=T7 promoter
                     /note="color: #ffffff"
     misc_feature    960..968
                     /label=Feature 13
                     /note="color: #a6acb3"
     misc_feature    963..968
                     /label=Kozak sequence
                     /note="color: #ff6600"
     misc_feature    969..2453
                     /label=Nier1s
                     /note="color: #a6acb3"
     misc_structure  969..1478
                     /label=mIFP1-170
                     /note="color: #ff99cc"
     misc_structure  1488..1928
                     /label=Calmodulin
                     /note="color: #ffcc00"
     misc_structure  1944..2012
                     /label=RS20
                     /note="color: #cc99ff"
     misc_structure  2025..2453
                     /label=mIFP178-320
                     /note="color: #ff99cc"
     CDS             2460..2480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             2484..2504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             2508..2528
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    2529..2594
                     /label=P2A
                     /note="Geneious type: motif"
                     /note="color: #a6acb3"
     primer_bind     2529..2547
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    2581..2595
                     /label=Gibson
                     /note="color: #a6acb3"
     primer_bind     2585..2600
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    join(2601..2795,2805..3317)
                     /label=eGFP
                     /label=eGFP(1)
                     /note="This feature has 2 segments:
                        1: 2601 .. 2795 / #00ff40
                        2: 2805 .. 3317 / #00ff40"
     misc_feature    2796..2804
                     /label=eGFP
                     /note="color: #00ff40"
     misc_feature    2796..2804
                     /label=eGFP Chromophore
                     /note="color: #800040"
     CDS             3324..3344
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             3348..3368
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             3372..3392
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    3393..3395
                     /label=Stop
                     /note="color: #ff0000"
     misc_feature    3413..3418
                     /label=AflII
                     /note="color: #008000"
     primer_bind     3436..3453
                     /label=BGH reverse primer
                     /note="color: #a6acb3; direction: RIGHT"
     polyA_site      3442..3666
                     /label=BGH pA
                     /note="color: #a6acb3"
     rep_origin      3712..4140
                     /label=f1 origin
                     /note="color: #ffff00"
     promoter        4145..4514
                     /label=SV40 early promoter
                     /note="color: #ffffff"
     CDS             4550..5344
                     /codon_start=1
                     /label=Neo
                     /label=Neo(R)
                     /note="color: #993366"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5518..5648
                     /label=SV40 pA
                     /note="color: #a6acb3"
     rep_origin      complement(6031..6701)
                     /direction=LEFT
                     /label=pUC origin
                     /note="color: #ffff00"
     CDS             complement(6846..7706)
                     /codon_start=1
                     /label=Amp
                     /label=Amp(R)
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7707..7805)
                     /label=bla promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggggcaaaga attggggttt
      901 gtctggtcaa ccaccgcggt ctccgtcgtc aggatcatcc gcagattata ccgcaactac
      961 acgccaccat gtcggtaccg ctgactacct cagcattcgg ccacgcgttt ctggctaact
     1021 gtgaacgtga gcagatccac ctggcgggct ccattcagcc gcacggtatc ctgctggctg
     1081 ttaaagagcc tgacaacgtg gtgatccagg cttctattaa cgctgcggag ttcctgaaca
     1141 ccaactttgt tgttggccgt ccgctgcgtg acctgggcgg cgatctgcct ttgcagatcc
     1201 tgccgcacct gaacggcccg ctgcacctgg ctccgatgac cctgcgttgt actgtgggtt
     1261 ctccgccgcg tcgtgtggac tgtaccattc atcgaccgtc taacggcggc ctgatcgtag
     1321 aactggaacc agcaaccaag gccactaaca ttgcgccggc tctgcagggt gcgcttcatc
     1381 gtatcacttc ttcatcctcc ctgatgggcc tgtgtgacga aaccgcgact attttccgtg
     1441 agattactgg cttcgaccgt gtgatggtaa tgcgtctcgg cgcgcttgac gatctgactg
     1501 aagagcagat cgcagagatt aaagaggctt tctccctatt tgacaaggac ggggacggga
     1561 cgataacaac caaggagctg gggacggtgt tccggtctct ggggcagaac cccacagaag
     1621 cagagctgca ggacatgatc aatgaagtag atgccgatgg cgacggcatc ttcgacttcc
     1681 ctgagttcct gacgatgatg gcaaggaaaa tgaatgactc agacagtgaa gaggaaatta
     1741 gaggcgcgtt ccgcgtgttt gataaggacg gcaatggcta catcggcgca gcagagcttc
     1801 gccacgtgat gacagacctt ggtgagaagt taactgatga ggaggttgat gaaatgatca
     1861 gggtagcaga caaggatggg gatggtcagg taaactacga agagtttgta caaatgatga
     1921 cagcgaaggg tggcggaggt tctgtagatt catcacgtcg taagtggaat aaggcaggtc
     1981 acgcagtcag agctataggt cggctgggct cgcgtaggca tgatttgctg tccgaatgtc
     2041 gtcgtgcgga cctggaagcg ttcctgggta accgctaccc ggcgtctact attccgcaga
     2101 tcgctcgtcg cctgtacgaa cttaaccgtg ttcgcctgct ggtagatgtg aactatactc
     2161 cggttccgct agagccgcgc atcagcccgc tgaacggtcg tgatctggat atgtccctgt
     2221 cttgcctgcg ctctatgtcc ccgatccacc agaaatacat gcaggacatg ggcgttggcg
     2281 caaccctggt ttgctctctg atggtgtctg gtcgtctgtg gggtctgatc gtttgccacc
     2341 actacgaacc gcgctacgtt ccgatgcaca ttcgcgctgc tggcgaagcg ctggcggaag
     2401 cctgtgcgaa ccgcatcgcg acgctggaga gctttgcaca gtctcagtcc aaaaccaggc
     2461 cgaaaaagaa gcgcaaggtt gaccctaaga agaagcgtaa agtcgaccca aagaagaaac
     2521 ggaaggtggg ctctggagcc accaacttca gcctgctgaa gcaggcaggc gacgtggaag
     2581 agaaccctgg ccctggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2641 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2701 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2761 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2821 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2881 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2941 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3001 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3061 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3121 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3181 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3241 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3301 tggacgagct gtacaagacg cgtccaaaaa agaagagaaa ggtagacccg aagaaaaagc
     3361 gtaaggtcga tcccaaaaag aaacggaaag tgtaagttta aatggtctcc agcttaagtt
     3421 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3481 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3541 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3601 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3661 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3721 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3781 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3841 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3901 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3961 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4021 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4081 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4141 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4201 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4261 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4321 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4381 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4441 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4501 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4561 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4621 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4681 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4741 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4801 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4861 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4921 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4981 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5041 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5101 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5161 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5221 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5281 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5341 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5401 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5461 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5521 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5581 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5641 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5701 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5761 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5881 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6601 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     6661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7741 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7801 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
