LOCUS       Exported                7389 bp DNA     circular SYN 11-OCT-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7389)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 11, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             907..939
                     /codon_start=1
                     /label=Lyn Targeting
                     /note="color: #800080"
                     /translation="MGCIKSKRKDK"
     regulatory      946..955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             952..1641
                     /codon_start=1
                     /label=ddGFP-A
                     /note="color: #008f00"
                     /translation="MVSKSEEVIKEFMRFKVRLEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPLIMYGSKMYVKHPADVPDYMKLSFPEGFKWERVMHFEDGG
                     LVTATQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTLGWDYATERLYPEEGVLKGELL
                     GRLKLKDGGLNLVESKTIYMAKKPVQLPGYYFVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLGHGTGS"
     CDS             1648..2106
                     /codon_start=1
                     /label=PH-PLCd1
                     /note="color: #803f02"
                     /translation="MDSGRDFLTLHGLQDDPDLQALLKGSQLLKVKSSSWRRERFYKLQ
                     EDCKTIWQESRKVMRSPESQLFSIEDIQEVRMGHRTEGLEKFARDIPEDRCFSIVFKDQ
                     RNTLDLIAPSPADAQHWVQGLRKIIHHSGSMDQRQKLQHWIHSCLRKAD"
     CDS             2113..2172
                     /codon_start=1
                     /label=PHdomain-Pseudoligand
                     /note="color: #84a4c0"
                     /translation="GGSVAEEEDDEEEDEDDGGS"
     CDS             2179..2877
                     /codon_start=1
                     /label=FP-B
                     /note="color: #800000"
                     /translation="ASSEDNNMATIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSEAYVRHPADIPDYKKLPFPEGFKWERVMNFED
                     GGLVTVTQDSSLQDGTLICKVKMRGTNFPPDGPVMQKKTMGWEASTEMLYPEDGVLKGH
                     SYQALKLKDGGHYLVEFETIYMAKKPVQLPGDYCVDTKLDITSHNEDYTIVEQYGRSEG
                     RHRLGMDERYK"
     promoter        complement(2941..2959)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    2985..3209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3255..3683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3697..4027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3878..4013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4094..4888
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5062..5183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5232..5248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5256..5272
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(5280..5310)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5280 .. 5286 / #ffffff / -10
                        2: 5287 .. 5304 / #ffffff
                        3: 5305 .. 5310 / #ffffff / -35"
     protein_bind    5325..5346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5634..6222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6393..7253)
                     /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: 6393 .. 7184 / #ccffcc
                        2: 7185 .. 7253 / #ccffcc / signal sequence
                     Cleavage site after base 7184"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7254..7358)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggagacccaa gcttgcggcc
      901 gccaccatgg gctgcatcaa gagcaagcgc aaggacaagg atccggccac catggtgagc
      961 aagagcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcttgga gggctccatg
     1021 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccttacga gggcacccag
     1081 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     1141 cccctgatca tgtacggctc caagatgtac gtgaagcacc ccgccgacgt ccccgattac
     1201 atgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgcactt cgaggacggc
     1261 ggtctggtga ccgcgacaca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     1321 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccttgggc
     1381 tgggattatg ccaccgagcg cctgtacccc gaagaaggcg tgctgaaggg cgagctcctg
     1441 gggcgcctga agctgaagga cggcggcctc aacctggtgg agtccaagac catctacatg
     1501 gccaagaagc ccgtgcaact gcccggctac tacttcgtgg acaccaagct ggacatcacc
     1561 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     1621 ctggggcatg gcaccggcag cgcatgcatg gactcgggta gggacttcct gaccctgcac
     1681 gggctccagg atgacccgga ccttcaggcc cttctgaagg gcagccagct tctgaaggtg
     1741 aagtccagct cgtggcgtag ggaacgcttc tacaagctac aggaggactg caagaccatc
     1801 tggcaggaat ctcgaaaggt catgaggtcc ccggagtcgc agctgttctc catcgaggac
     1861 attcaggagg tacggatggg acaccgcaca gaaggcctgg agaagtttgc ccgagacatc
     1921 cccgaggatc gatgcttctc cattgtcttc aaggaccagc gcaacaccct agacctcatt
     1981 gccccatcac cagctgacgc tcagcactgg gtgcagggcc tgcgcaagat catccaccac
     2041 tccggctcca tggaccagcg gcagaagctg cagcactgga ttcactcctg cttgcgaaag
     2101 gctgatgtcg acggcggtag cgtggcagaa gaggaagatg acgaggaaga ggacgaggac
     2161 gatggcggca gcgagctcgc ctcctccgag gacaacaaca tggccaccat caaagagttc
     2221 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     2281 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     2341 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccgaggcg
     2401 tacgtgaggc accccgccga catccccgat tacaagaagc tgcccttccc cgagggcttc
     2461 aagtgggagc gcgtgatgaa cttcgaagac ggcggtctgg tgaccgttac ccaggactcc
     2521 tccctgcagg acggcacgct gatctgcaag gtgaagatgc gcggcaccaa cttccccccc
     2581 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gatgctgtac
     2641 cccgaagacg gcgtgctgaa gggccatagc tatcaggccc tgaagctgaa ggacggcggc
     2701 cactacctgg tggagttcga gaccatctac atggccaaga agcccgtgca actgcccggc
     2761 gattactgtg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     2821 gaacagtacg ggcgctccga gggccgccac cgtctgggca tggacgagcg gtacaagtaa
     2881 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat
     2941 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3001 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3061 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3121 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3181 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3241 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3301 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3541 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3601 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3661 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3721 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3901 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3961 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4021 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4081 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4141 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4201 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4261 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4321 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4381 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4441 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4501 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4561 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4621 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4681 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4741 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4801 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4861 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4921 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4981 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5041 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5161 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5221 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5821 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6061 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7381 cctgacgtc
//
