LOCUS       Exported                7422 bp DNA     circular SYN 11-OCT-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 11, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
COMMENT     Ligation of : pcDNA3-AKAR4.gb.xdna [5348  nt] : (XbaI[3031] / 
            HindIII[889]) to pcDNA3-Lyn-PIP3TIR-ddRFP.gb.xdna [2066  n t] : 
            (HindIII[1] / XbaI[2071])
FEATURES             Location/Qualifiers
     source          1..7422
                     /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=ddRFP-A
                     /note="color: #ff0000"
                     /translation="MVSKSEEVIKEFMRFKVRLEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADVPDYMKLSFPEGFKWERVMHFEDGG
                     LVTVTQDTSLQDGTLIYKVKMRGTNFPPDGPVMQKKTLGWDYATERLYPEDGVLKGELL
                     GRLKLKDGGLNLVEFKTIYMAKKPVQLPGYYFVDTKLGITSHNEDYTIVEQYERSEGRH
                     HLGHGTGS"
     CDS             1648..2139
                     /codon_start=1
                     /label=PH-Akt(R23/25A)
                     /note="color: #803f02"
                     /translation="MNDVAIVKEGWLHKRGEYIKTWAPAYFLLKNDGTFIGYKERPQDV
                     EQRESPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKRQEEETMDFRSGSPSDNSGAEEMEVALAKPKHRVTMNEFEYLKLLGKGTFGK
                     V"
     CDS             2146..2205
                     /codon_start=1
                     /label=PHdomain-Pseudoligand
                     /note="color: #84a4c0"
                     /translation="GGSVAEEEDDEEEDEDDGGS"
     CDS             2212..2910
                     /codon_start=1
                     /label=FP-B
                     /note="color: #800000"
                     /translation="ASSEDNNMATIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSEAYVRHPADIPDYKKLPFPEGFKWERVMNFED
                     GGLVTVTQDSSLQDGTLICKVKMRGTNFPPDGPVMQKKTMGWEASTEMLYPEDGVLKGH
                     SYQALKLKDGGHYLVEFETIYMAKKPVQLPGDYCVDTKLDITSHNEDYTIVEQYGRSEG
                     RHRLGMDERYK"
     promoter        complement(2974..2992)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    3018..3242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3288..3716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3730..4060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4127..4921
                     /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    5095..5216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5265..5281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5289..5305
                     /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(5313..5343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5313 .. 5319 / #ffffff / -10
                        2: 5320 .. 5337 / #ffffff
                        3: 5338 .. 5343 / #ffffff / -35"
     protein_bind    5358..5379
                     /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(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6426..7286)
                     /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: 6426 .. 7217 / #ccffcc
                        2: 7218 .. 7286 / #ccffcc / signal sequence
                     Cleavage site after base 7217"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7287..7391)
                     /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 gcccctacga gggcacccag
     1081 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     1141 ccccagatca tgtacggctc caaggcgtac gtgaagcacc ccgccgacgt ccccgattac
     1201 atgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgcactt cgaggacggc
     1261 ggtctggtga ccgtgactca ggacacctcc ctgcaggacg gcacgctgat ctacaaggtg
     1321 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccttgggc
     1381 tgggattatg ccaccgagcg cctgtacccc gaagacggcg tgctgaaggg cgagcttctg
     1441 gggcgcctga agctgaagga cggcggcctc aacctggtgg agttcaagac catctacatg
     1501 gccaagaagc ccgtgcaact gcccggctac tacttcgtgg acaccaagct gggcatcacc
     1561 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     1621 ctggggcatg gcaccggcag cgcatgcatg aacgacgtag ccattgtgaa ggagggctgg
     1681 ctgcacaaac gaggggaata tattaaaacc tgggcgccag cctacttcct cctcaagaat
     1741 gatggcacct ttattggcta caaggaacgg cctcaggatg tggagcagcg cgagtcccca
     1801 ctcaacaact tctcagtggc acaatgtcag ctgatgaaga cagagcggcc gaggcccaac
     1861 accttcatca tccgctgcct gcagtggacc acggtcattg agcgcacctt ccatgtggaa
     1921 acgcctgagg agcgggaaga gtggaccacc gccattcaga ctgtggctga tggactcaaa
     1981 cggcaggagg aggagacgat ggacttccgg tcaggttcac ccagtgacaa ctcaggtgct
     2041 gaggagatgg aggtggccct ggccaagccc aagcaccgtg tgaccatgaa cgagtttgag
     2101 tacctgaagc tactgggcaa gggcaccttt gggaaggtgg tcgacggcgg tagcgtggca
     2161 gaagaggaag atgacgagga agaggacgag gacgatggcg gcagcgagct cgcctcctcc
     2221 gaggacaaca acatggccac catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     2281 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2341 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     2401 ctgtcccccc agttcatgta cggctccgag gcgtacgtga ggcaccccgc cgacatcccc
     2461 gattacaaga agctgccctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgaa
     2521 gacggcggtc tggtgaccgt tacccaggac tcctccctgc aggacggcac gctgatctgc
     2581 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     2641 atgggctggg aggcctccac cgagatgctg taccccgaag acggcgtgct gaagggccat
     2701 agctatcagg ccctgaagct gaaggacggc ggccactacc tggtggagtt cgagaccatc
     2761 tacatggcca agaagcccgt gcaactgccc ggcgattact gtgtggacac caagctggac
     2821 atcacctccc acaacgagga ctacaccatc gtggaacagt acgggcgctc cgagggccgc
     2881 caccgtctgg gcatggacga gcggtacaag taagaattct gcagatatcc atcacactgg
     2941 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3001 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3061 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3121 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3181 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3241 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3721 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3841 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3961 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4081 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4141 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4201 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4261 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4321 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4381 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4441 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4501 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4561 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4621 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4681 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4741 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4801 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4861 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4921 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4981 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5041 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5101 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5161 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5221 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5281 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5341 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5401 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5461 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
