LOCUS       G2_ssCAG-NLS-eGF        5419 bp    DNA     circular SYN 30-OCT-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
COMMENT     LOCUS       1787\-\pAAVio-CA        4902 bp    DNA     circular SYN 
            31-AUG-2011

            DEFINITION  plasmid HIV Alex George Enhanced.

            ACCESSION   Your mama'

            SOURCE      Unknown.

              ORGANISM  Unknown

                        Unclassified.

            REFERENCE   1  (bases 1 to 8541)

              AUTHORS   Self

              JOURNAL   Unpublished.

            COMMENT     HRST Ligated into PUC

                        WPRE fixed

                        Changed cPPT to include SpeI site for cloning promoters

                        cloned in clean CMV promoter without extraneous 
            sequence

                        9/19/03 Completely sequenced with 4x coverage to confirm
            perfect sequence

                        11/13/03 Confirmed shortened cloning to add SmaI/PacI 
            and tag

                        11/20/03 Tested Virus production and exression from 
            viral integration in parallell with all Intermediate Vectors and 
            found good titer and expression

                        1/14/04  Confirmed Insulator insertion to have perfect 
            sequence

                        3/18/04 Confirmed perfect recreation of 3' LTR and W to 
            remove extraneous chromosomal sequence, add directional 8bp cloning 
            sites to dU3 and shorten it to the same length as pHAGEv1.0

                        4/1/04 Confirmed perfect 5' LTR sequence containing 
            major Splice Donor Site

                        5/5/04 Confirmed perfect SV40 to Xho Sequence, CAG-R 
            fusion was also done perfectly.  CAG Promoter appears to be lacking 
            54 bp in the beta-actin portion of the promoter, unclear if this is 
            a PCR error or template issue may still be a functional promoter

                        5/5/04 Confirmed perfect cPPT fixed sequence

                        6/18/04 Sequencing confirmed perfect CMV-R Fusion

                        11/06/05 Sequencing confirms perfect replacement of SV40
            with linker sequence

                        03/08/06 Sequencing confirms perfect Beta-Globin Intron 
            with cloned into PSI Region with Mutant Splice Donor

            COMMENT     This file is created by Vector NTI

                        http://www.invitrogen.com/

            COMMENT     ORIGDB|GenBank

            COMMENT     VNTDATE|599931188|

            COMMENT     VNTDBDATE|599931188|

            COMMENT     LSOWNER|

            COMMENT     VNTNAME|1787 - pAAVio-CASI-eGFP-W-SV40|

            COMMENT     VNTAUTHORNAME|Alejandro Balazs|

            COMMENT     VNTAUTHOREML|alexb@caltech.edu|
FEATURES             Location/Qualifiers
     source          1..5419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3509..3945
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     rep_origin      1..534
                     /direction=RIGHT
                     /label=ORI
                     /note="Bacterial Origin of Replication"
                     /note="color: #c0c0c0"
     primer_bind     206..230
                     /label=pHAGE-S7501-8000
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     complement(473..494)
                     /label=pHAGE-A7550-7050
                     /note="color: #ff8080; direction: LEFT"
     LTR             653..775
                     /label=ITR1
                     /note="color: #ffff00; direction: BOTH"
     misc_feature    696..712
                     /label=Flip
                     /note="color: #ff8000"
     misc_feature    777..1881
                     /label=Untitled
                     /note="color: #000000"
     promoter        781..1880
                     /label=CAG
                     /note="CAG"
                     /note="color: #00ffe1; direction: RIGHT"
     primer_bind     1858..1943
                     /label=CTCATCATTTTGGCAAAGAATTGCGGCCGCCACCATGGGAAGCCCAAAGA
                     /note="color: black; sequence: 
                     CTCATCATTTTGGCAAAGAATTGCGGCCGCCACCATGGGAAGCCCAAAGAAGAAGCGTA
                     AGGTAATGGTGAGCAAGGGCGAGGAGC"
     misc_feature    1886..1895
                     /label=RBS - ribosome  binding site
                     /note="RBS - ribosome  binding site"
                     /note="color: #ff8000"
     misc_feature    1895..1900
                     /label=GS extra residues
                     /note="GS extra residues"
                     /note="color: #ff00fe"
     misc_feature    1901..1921
                     /label=NLS
                     /note="NLS"
                     /note="color: #ff8000"
     CDS             1922..2638
                     /label=eGFP
                     /note="color: #0000ff; direction: RIGHT"
     primer_bind     2011..2028
                     /label=pHAGE-S3001-3500
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     complement(2254..2274)
                     /label=pHAGE-A3050-2550
                     /note="color: #ff8080; direction: LEFT"
     primer_bind     2509..2530
                     /label=pHAGE-S3501-4000
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     complement(2621..2679)
                     /label=CTTGTACAGCTCGTCCAT
                     /note="color: black; sequence: 
                     GATTAGGATCCTTACACCTTCCGCTTTTTCTTGGGGCTTCCCTTGTACAGCTCGTCCAT
                     "
     misc_feature    2639..2644
                     /label=GS extra residues
                     /note="GS extra residues"
                     /note="color: #ff00fe"
     misc_feature    2645..2664
                     /label=NLS
                     /note="NLS"
                     /note="color: #ff8000"
     misc_feature    2670..2673
                     /label=Untitled
                     /note="color: #------"
     misc_feature    2675..3266
                     /label=WPRE
                     /note="color: #ff8000"
     primer_bind     complement(2703..2737)
                     /label=pHAGE 3' CDS
                     /note="color: #ff8080; direction: LEFT"
     primer_bind     complement(2781..2804)
                     /label=pHAGE-A3550-3050
                     /note="color: #ff8080; direction: LEFT"
     primer_bind     2934..2954
                     /label=pHAGE-S4001-4500
                     /note="color: #ff8080; direction: RIGHT"
     misc_feature    3131..3150
                     /label=Untitled
                     /note="color: #ff8000"
     polyA_signal    3273..3505
                     /label=SV40 late polyadenylation signal
                     /note="color: #8000ff"
     misc_feature    3398..3403
                     /label=Untitled
                     /note="color: #ff8000"
     misc_feature    3509..3945
                     /label=Zombie Construct
                     /note="color: #ff0000"
     misc_feature    3509..3891
                     /label=Barcode 6
                     /note="color: #a6acb3"
     misc_feature    3516..3540
                     /label=P14
                     /note="color: #a6acb3"
     misc_feature    3543..3567
                     /label=P13
                     /note="color: #a6acb3"
     misc_feature    3576..3595
                     /label=SNP-P10
                     /note="color: #a6acb3"
     misc_feature    3631..3650
                     /label=SNP-P9
                     /note="color: #a6acb3"
     misc_feature    3658..3677
                     /label=SNP-P8
                     /note="color: #a6acb3"
     misc_feature    3686..3705
                     /label=SNP-P7
                     /note="color: #a6acb3"
     misc_feature    3718..3737
                     /label=SNP-P6
                     /note="color: #a6acb3"
     misc_feature    3748..3767
                     /label=SNP-P5
                     /note="color: #a6acb3"
     misc_feature    3788..3807
                     /label=SNP-P4
                     /note="color: #a6acb3"
     misc_feature    3818..3837
                     /label=SNP-P3
                     /note="color: #a6acb3"
     misc_feature    3840..3864
                     /label=P2
                     /note="color: #a6acb3"
     misc_feature    3840..3859
                     /label=SNP-P2
                     /note="color: #a6acb3"
     misc_feature    3867..3891
                     /label=P1
                     /note="color: #a6acb3"
     misc_feature    complement(3892..3945)
                     /label=promoter
                     /note="color: #3366ff; direction: LEFT"
     promoter        complement(3893..3911)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(3922..3940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     LTR             complement(3949..4093)
                     /label=ITR2
                     /note="color: #ffff00; direction: LEFT"
     misc_feature    4012..4050
                     /label=Flop
                     /note="color: #ff8000"
     primer_bind     4128..4150
                     /label=pHAGE-S6001-6500
                     /note="color: #ff8080; direction: RIGHT"
     promoter        4299..4333
                     /label=P-Bla
                     /note="Beta Lactamase Promoter"
                     /note="color: #8080ff"
     CDS             4352..5191
                     /label=AMPr
                     /note="Ampicillin Resistance Gene"
                     /note="color: #0000ff; direction: RIGHT"
     primer_bind     complement(4401..4424)
                     /label=pHAGE-A6050-5550
                     /note="color: #ff8080; direction: LEFT"
     primer_bind     4601..4625
                     /label=pHAGE-S6501-7000
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     complement(4870..4893)
                     /label=pHAGE-A6550-6050
                     /note="color: #ff8080; direction: LEFT"
     primer_bind     5121..5145
                     /label=pHAGE-S7001-7500
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     complement(5381..5402)
                     /label=pHAGE-A7050-6550
                     /note="color: #ff8080; direction: LEFT"
ORIGIN
        1 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
       61 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtaataa
      601 acacacacac accaacaacc gtggttggtt gttgtgttgg tttattctcg agttggccac
      661 tccctctctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcctca
      721 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctactag
      781 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      841 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      901 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      961 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1021 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1081 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1141 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     1201 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     1261 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
     1321 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     1381 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     1441 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1501 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     1561 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     1621 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
     1681 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
     1741 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
     1801 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1861 atcattttgg caaagaattg cggccgccac catgggaagc ccaaagaaga agcgtaaggt
     1921 aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1981 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2041 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2101 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2161 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2221 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2281 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2341 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2401 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2461 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2521 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2581 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     2641 aagccccaag aaaaagcgga aggtgtaagg atcctaatca acctctggat tacaaaattt
     2701 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2761 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2821 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2881 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2941 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3001 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3061 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3121 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3181 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3241 tctccctttg ggccgcctcc ccgcctggta ccttcgagca gacatgataa gatacattga
     3301 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3361 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3421 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3481 aaacctctac aaatgtggta aaatcaagcg acagcttgtc tctccagatg ctcttgggcc
     3541 atcttccaca tcgtccgtag cagccttggc aatttgccat cactggcaaa tacacataaa
     3601 tccaatgaat acggttacca ccatcacatt accatgcagg tacacagcaa gaattgacgt
     3661 tggcatatca catggtgtaa taaccccact tgtgaaacaa cccagaataa ggtacaaggc
     3721 ggaaatgtcg tcattctaaa ataaaaggca tggccaggaa tttgtctaat accgggaact
     3781 taaattcagc ttgaacacca gtcgcaaaaa attcaaagaa agtgattcag gttcgggttc
     3841 gtggattgga acagcttctt ttgtttcagt gatgagagaa tcctcctgtc accctttagt
     3901 gagggttaat tctcgagtct ccctatagtg agtcgtatta attcccttag gaacccctag
     3961 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4021 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag
     4081 agggagtggc caagctagcg ggcgattaag gaaagggcta gatcattctt gaagacgaaa
     4141 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4201 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4261 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4321 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4381 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4441 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4501 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4561 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4621 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4681 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4741 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4801 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4861 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4921 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4981 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5041 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5101 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5161 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5401 gcaaacaaaa aaaccaccg
//
