LOCUS       pBEL3002                6926 bp ds-DNA     circular     06-MAR-2024
DEFINITION  .
KEYWORDS    "Key Features:40 amino acids of SP(Eint_071040)-sfGFP, secreted?,
            initial 40 residues of Eint_071040/PTP5" "Backbone:pBEL1764"
            "Construction:Gibson cloning of 40 amino acids of
            SP(Eint_071040)-sfGFP into EcoRI-NotI-digested pBEL1764."
            "Resistance:Carb" "Location" "Rack" "Box" "Notes" "Sequencing
            information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pcDNA3.4-TOPO
            (linearized).gb Linearized mammalian vector with 3'-T overhangs and
            bound topoisomerase, for TOPO(R) TA cloning of PCR products and
            high level expression. Do not use phosphorylated primers for the
            PCR amplification because 5' phosphates will prevent ligation into
            this vector.
FEATURES             Location/Qualifiers
     enhancer        47..426
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          341..360
                     /label="oDCE3730"
                     /note="sequence: ccgcctggcattatgcccag"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     promoter        427..630
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          581..601
                     /label="CMV-Forward"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          637..657
                     /label="oDCE3706"
                     /note="sequence: gtgaaccgtcagatcgcctgg"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    739..762
                     /label="Additional 5' UTR from Rb IgH +control plasmid"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    763..777
                     /label="5' UTR Rb IgH mRNA"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    778..780
                     /label="START"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             778..1611
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MILILVLGMAFSYPLYLDSFTNKPIRILAAEYHLLVSELKSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK*"
     misc_feature    781..897
                     /label="SP, E0S832"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(819..926)
                     /label="oCL019"
                     /note="sequence: ACTCCAGTGAAAAGTTCTTCTCCTTTGCTTTTGAGTTCGGAGACAAGAAGATGATACTCTGCTGCCAAGATTCTGATAGGTTTGTTGGTAAATGAGTCGAGGTATAGC"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    898..1608
                     /label="sfGFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(1583..1657)
                     /label="oDCE4020"
                     /note="sequence: cttGTCGAGGTCGGGGGATCCGGCCTTGCCGGCCTCGAGCGGCCGCTCATTTGTAGAGCTCATCCATGCCATGTG"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    1609..1611
                     /label="STOP"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1612..1654
                     /label="Additional 3' UTR from Rb IgH +control plasmid"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1700..2288
                     /label="WPRE"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(1767..1788)
                     /label="oDCE3707"
                     /note="sequence: aaagcagcgtatccacatagcg"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          2148..2168
                     /label="oDCE3709"
                     /note="sequence: ggacgtccttctgctacgtcc"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(2326..2345)
                     /label="oDCE3374"
                     /note="sequence: GCGGGTTCCTTCCGGTATTG"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     polyA_signal    2353..2401
                     /label="HSV TK poly(A) signal"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      2603..3031
                     /label="f1 ori"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(2641..2659)
                     /label="oDCE3639"
                     /note="sequence: tcacgctgcgcgtaaccac"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          2663..2683
                     /label="oDCE3710"
                     /note="sequence: ctacacttgccagcgccctag"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        3045..3374
                     /label="SV40 promoter"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          3185..3209
                     /label="oDCE3711"
                     /note="sequence: caattagtcagcaaccatagtcccg"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     rep_origin      3225..3360
                     /label="SV40 ori"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             3441..4235
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          3781..3802
                     /label="oDCE3712"
                     /note="sequence: ccatcatggctgatgcaatgcg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(3864..3882)
                     /label="oDCE3446"
                     /note="sequence: TCCGAGTACGTGCTCGCTC"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          4384..4404
                     /label="oDCE3713"
                     /note="sequence: ctcatgctggagttcttcgcc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4411..4532
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(4448..4467)
                     /label="SV40pA-R"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(4595..4614)
                     /label="M13-48REV"
                     /note="sequence: CGGATAACAATTTCACACAG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          4975..4996
                     /label="oDCE3449"
                     /note="sequence: CTGGCGTTTTTCCATAGGCTCC"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     rep_origin      complement(4983..5571)
                     /label="ori"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(5180..5201)
                     /label="oDCE3562"
                     /note="sequence: ctgagatacctacagcgtgagc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(5287..5305)
                     /label="oDCE3451"
                     /note="sequence: CCGGGTTGGACTCAAGACG"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          5591..5610
                     /label="oDCE3548"
                     /note="sequence: ctacggggtctgacgctcag"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             complement(5742..6602)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(5795..5816)
                     /label="oDCE3561"
                     /note="sequence: gggagtcaggcaactatggatg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          6194..6213
                     /label="oDCE3547"
                     /note="sequence: gaagtaagttggccgcagtg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(6415..6435)
                     /label="oDCE3560"
                     /note="sequence: gagttttcgccccgaagaacg"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(6603..6707)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          6791..6813
                     /label="oDCE3714"
                     /note="sequence: ctctgatgccgcatagttaagcc"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
ORIGIN
        1 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
       61 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      661 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      721 ctagaggatc gaacccttGG ATCTCTAGCG AATTCCCTCT AGACACAGAC GCTCACCATG
      781 ATTCTGATAT TGGTTCTTGG AATGGCATTT TCCTATCCGC TATACCTCGA CTCATTTACC
      841 AACAAACCTA TCAGAATCTT GGCAGCAGAG TATCATCTTC TTGTCTCCGA ACTCAAAAGC
      901 AAAGGAGAAG AACTTTTCAC TGGAGTTGTC CCAATTCTTG TTGAATTAGA TGGTGATGTT
      961 AATGGGCACA AATTTTCTGT CCGTGGAGAG GGTGAAGGTG ATGCTACAAA CGGAAAACTC
     1021 ACCCTTAAAT TTATTTGCAC TACTGGAAAA CTACCTGTTC CGTGGCCAAC ACTTGTCACT
     1081 ACTCTGACCT ATGGTGTTCA ATGCTTTTCC CGTTATCCGG ATCACATGAA ACGGCATGAC
     1141 TTTTTCAAGA GTGCCATGCC CGAAGGTTAT GTACAGGAAC GCACTATATC TTTCAAAGAT
     1201 GACGGGACCT ACAAGACGCG TGCTGAAGTC AAGTTTGAAG GTGATACCCT TGTTAATCGT
     1261 ATCGAGTTAA AGGGTATTGA TTTTAAAGAA GATGGAAACA TTCTTGGACA CAAACTCGAG
     1321 TACAACTTTA ACTCACACAA TGTATACATC ACGGCAGACA AACAAAAGAA TGGAATCAAA
     1381 GCTAACTTCA AAATTCGCCA CAACGTTGAA GATGGTTCCG TTCAACTAGC AGACCATTAT
     1441 CAACAAAATA CTCCAATTGG CGATGGCCCT GTCCTTTTAC CAGACAACCA TTACCTGTCG
     1501 ACACAATCTG TCCTTTCGAA AGATCCCAAC GAAAAGCGTG ACCACATGGT CCTTCTTGAG
     1561 TTTGTAACTG CTGCTGGGAT TACACATGGC ATGGATGAGC TCTACAAATG AGCGGCCGCT
     1621 CGAGGCCGGC AAGGCCGGAT CCCCCGACCT CGACaagggt tcgatcccta ccggttagta
     1681 atgagtttga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1741 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1801 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1861 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1921 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     1981 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2041 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2101 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2161 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2221 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2281 ctccccgcct ggaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2341 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2401 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2461 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2521 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     2581 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2641 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2701 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2761 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2821 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2881 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2941 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3001 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3301 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3361 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3601 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3961 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4261 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4321 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4381 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4441 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4501 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4561 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4621 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5401 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6721 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     6781 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     6841 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     6901 attgcatgaa gaatctgctt agggtt
//