LOCUS       Exported                3435 bp ds-DNA     circular SYN 17-AUG-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3435)
  AUTHORS   Cohn Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 17, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3435
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..28
                     /label=WT-KnockIn-F
                     /note="2746 64.249976389621"
                     /note="color: black; sequence: 
                     GACGTCGCTAGCTCTAGAGGGCCTGAAT; added: 2020-10-13"
     primer_bind     1..22
                     /label=puro-Ins_IF_F
                     /note="4353 70.2057831440608"
                     /note="color: black; sequence: 
                     ACTGGATCCGGTACCGACGTCGCTAGCTCTAGAGGGC; added: 2020-10-13"
     primer_bind     complement(7..29)
                     /label=Linear BPK_MCS_R
                     /note="2273 58.227245756756"
                     /note="color: black; sequence: AATTCAGGCCCTCTAGAGCTAGC; 
                     added: 2020-10-13"
     primer_bind     complement(7..24)
                     /label=roundTwoQ5-R
                     /note="2716 55.6657288297645"
                     /note="color: black; sequence: AGGCCCTCTAGAGCTAGC; added: 
                     2020-10-13"
     primer_bind     8..30
                     /label=BPK1520_MCS_F
                     /note="2220 56.3722255693635"
                     /note="color: black; sequence: CTAGCTCTAGAGGGCCTGAATTC; 
                     added: 2020-10-13"
     primer_bind     complement(10..26)
                     /label=knockIn-Q5-round2-CORRECT-R
                     /note="2718 60.9848250024049"
                     /note="color: black; sequence: 
                     TTGATATACAAATAGGTCAGGCCCTCTAGAGCT; added: 2020-10-13"
     primer_bind     complement(12..30)
                     /label=BPK1520_MCS_R
                     /note="color: black; sequence: gatcgaattcaggccctctagag"
     primer_bind     complement(12..30)
                     /label=BPK1520_MCS_Ra
                     /note="2221 56.5686762323769"
                     /note="color: black; sequence: GATCGAATTCAGGCCCTCTAGAG; 
                     added: 2020-10-13"
     primer_bind     15..48
                     /label=In-Fusion_puro_F
                     /note="color: black; sequence: 
                     TAGAGGGCCTGAATTCTCGAGGCGTTGACATTGA"
     primer_bind     30..47
                     /label=px601 seq1 fw
                     /note="479"
                     /note="color: black; sequence: GACTCGAGGCGTTGACATTG"
     primer_bind     33..70
                     /label=cas13b-inFusion-puroIns-F
                     /note="3796 66.4719089367442"
                     /note="color: black; sequence: 
                     TGACCGTCTCCGGGAGAGGCGTTGACATTGATTATTGACTAGTTATTAATAGT; 
                     added: 2020-10-13"
     primer_bind     complement(35..66)
                     /label=knockInCloning_R
                     /note="2668 55.8216063091629"
                     /note="color: black; sequence: 
                     TTAATAACTAGTCAATAATCAATGTCAACGCC; added: 2020-10-13"
     enhancer        41..420
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     primer_bind     41..83
                     /label=CMV-FKBP Fragment.FOR
                     /note="4011 64.58036508"
                     /note="color: black; sequence: 
                     GTCGGAGCAAGCTTGGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGG;
                      added: 2020-10-13"
     primer_bind     41..83
                     /label=CMV-Frb Fragment.FOR
                     /note="4015 64.58036508"
                     /note="color: black; sequence: 
                     GTCGGAGCAAGCTTGGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGG;
                      added: 2020-10-13"
     primer_bind     41..69
                     /label=CMV_enh_EcoRI-F
                     /note="2032 57.5908118314277"
                     /note="color: black; sequence: 
                     GCTCGAATTCAAGACATTGATTATTGACTAGTTATTAATAG; added: 
                     2020-10-13"
     primer_bind     complement(99..118)
                     /label=px601 seq3 rv
                     /note="508"
                     /note="color: black; sequence: CGCGGAACTCCATATATGGG"
     primer_bind     113..133
                     /label=CMV_enh_BamHI-F
                     /note="color: black; sequence: 
                     GAGTggaTCCGCGTTACATAACTTACGG"
     primer_bind     113..133
                     /label=CMV_enh_BamHI-Fa
                     /note="358 60.387849426302601"
                     /note="color: black; sequence: 
                     GAGTGGATCCGCGTTACATAACTTACGG; added: 2020-10-13"
     primer_bind     complement(398..420)
                     /label=CMV_enh_EcoRI-R
                     /note="2033 56.5577393913329"
                     /note="color: black; sequence: 
                     CAGTGAATTCATGGTAATAGCGATGACTAATA; added: 2020-10-13"
     promoter        421..624
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     521..544
                     /label=CMV_prom_F
                     /note="137"
                     /note="color: black; sequence: GGCACCAAAATCAACGGGACTTTC"
     primer_bind     537..554
                     /label=VP1.5
                     /note="1"
                     /note="color: black; sequence: GGACTTTCCAAAATGTCG"
     primer_bind     537..554
                     /label=VP1.5 (2)
                     /note="3"
                     /note="color: black; sequence: GGACTTTCCAAAATGTCG"
     primer_bind     complement(600..624)
                     /label=Vector.REV (2)
                     /note="2487 57.4506984393103"
                     /note="color: black; sequence: AGCTCTGCTTATATAGACCTCCCAC; 
                     added: 2020-10-13"
     primer_bind     601..620
                     /label=CMV Seq Primer
                     /note="1077"
                     /note="color: black; sequence: TGGGAGGTCTATATAAGCAG"
     primer_bind     604..624
                     /label=px601 seq2 fw
                     /note="480"
                     /note="color: black; sequence: GAGGTCTATATAAGCAGAGCT"
     primer_bind     complement(607..637)
                     /label=DNMT3A-vector-R
                     /note="2570 58.2415334636833"
                     /note="color: black; sequence: 
                     TAGTTAGCCAGAGAGCTCTGCTTATATAGAC; added: 2020-10-13"
     primer_bind     complement(607..637)
                     /label=hHDAC-vector-R
                     /note="2568 58.2415334636833"
                     /note="color: black; sequence: 
                     TAGTTAGCCAGAGAGCTCTGCTTATATAGAC; added: 2020-10-13"
     primer_bind     complement(625..642)
                     /label=Q5_kozak_puroR_R
                     /note="color: black; sequence: accggtagttagccagag"
     primer_bind     complement(625..642)
                     /label=Q5_kozak_puroR_Ra
                     /note="2325 53.4186802786958"
                     /note="color: black; sequence: ACCGGTAGTTAGCCAGAG; added: 
                     2020-10-13"
     primer_bind     complement(634..651)
                     /label=Vector.REV
                     /note="color: black; sequence: CATggtggcaccggtagt; added: 
                     2020-12-17"
     primer_bind     complement(635..652)
                     /label=CMV-FKBP Fragment.REV
                     /note="4012 65.12285927"
                     /note="color: black; sequence: 
                     ACAAGTAGCTTGTATCCATGGTGGCACCGGTAG; added: 2020-10-13"
     primer_bind     complement(635..652)
                     /label=CMV-Frb Fragment.REV
                     /note="4016 65.12285927"
                     /note="color: black; sequence: 
                     ACAAGTAGCTTGTATCCATGGTGGCACCGGTAG; added: 2020-10-13"
     primer_bind     637..670
                     /label=Fragment.FOR
                     /note="color: black; sequence: 
                     accggtgccaccATGgccatgcctttgtctcaag; added: 2020-12-17"
     regulatory      643..652
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             649..1045
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /note="color: #ccffcc"
                     /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(770..791)
                     /label=HAP1Cas9-3'-R2
                     /note="2749 55.5749473208204"
                     /note="color: black; sequence: ACATTGACACCAGTGAAGATGC; 
                     added: 2020-10-30"
     primer_bind     complement(1027..1059)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     atcgatgaattcTCAgccctcccacacataacc; added: 2020-12-17"
     primer_bind     1045..1070
                     /label=Vector.FOR
                     /note="color: black; sequence: TGAgaattcatcgataccgtcgagat; 
                     added: 2020-12-17"
     primer_bind     1047..1072
                     /label=linear Sacas9 VP64 C term F
                     /note="4923 56.913004278881"
                     /note="color: black; sequence: 
                     TAAGAATTCATCGATACCGTCGAGATCT; added: 2020-10-13"
     polyA_site      1079..1205
                     /label=SV40 Poly A Site
                     /note="color: #ffffff"
     primer_bind     1109..1133
                     /label=SV40-Fw
                     /note="1722"
                     /note="color: black; sequence: AGCAATAGCATCACAAATTTCACAA"
     primer_bind     complement(1129..1156)
                     /label=T385M Q5 LoxP Proximal F
                     /note="4535 59.165837993653"
                     /note="color: black; sequence: 
                     CATTATACGAAGTTATGCAGTGAAAAAAATGCTTTATTTGTG; added: 
                     2020-10-13"
     primer_bind     complement(1176..1204)
                     /label=SV40-Rev
                     /note="1723"
                     /note="color: black; sequence: 
                     CCAGACATGATAAGATACATTGATGAGTT"
     primer_bind     complement(1182..1220)
                     /label=In-Fusion_puro_R
                     /note="color: black; sequence: 
                     GTACCGGATCGAATTTCCAGACATGATAAGATACATTGA"
     primer_bind     complement(1183..1205)
                     /label=A5-InFusion-Vect-F
                     /note="2458 51.2763098992908"
                     /note="color: black; sequence: GATCCAGACATGATAAGATACATTG; 
                     added: 2020-10-13"
     primer_bind     complement(1199..1221)
                     /label=cas13b-inFusion-puroIns-R
                     /note="3797 68.0539445926696"
                     /note="color: black; sequence: 
                     CTTTCGTCTCGCGCGGTACCGGATCGAATTTCCAGAC; added: 2020-10-13"
     primer_bind     1206..1228
                     /label=Linear BPK_MCS_F
                     /note="2272 58.8801917398693"
                     /note="color: black; sequence: AATTCGATCCGGTACCAAGGTCG; 
                     added: 2020-10-13"
     primer_bind     1209..1229
                     /label=BPK2660-U6-F
                     /note="3074 66.4660188807203"
                     /note="color: black; sequence: 
                     ACCTCTACAAATGTGTCGATCCGGTACCAAGGTCGG; added: 2020-10-13"
     primer_bind     1211..1229
                     /label=KpnI_U6 fw
                     /note="514 67.599999999999994"
                     /note="color: black; sequence: GCGCGGATCCGGTACCAAGGTCGG; 
                     added: 2020-10-13"
     primer_bind     complement(1214..1234)
                     /label=puro-Ins_IF_R
                     /note="4354 71.0197053242316"
                     /note="color: black; sequence: 
                     TTCCTGCCCGACCTTCCCGACCTTGGTACCGG; added: 2020-10-13"
     primer_bind     complement(1219..1236)
                     /label=bpk1520-Nme2_VECT_R
                     /note="5320 59.3661196848116"
                     /note="color: black; sequence: TTCCTGCCCGACCTTGGT; added: 
                     2020-10-13"
     primer_bind     complement(1219..1236)
                     /label=bpk1520-peg-Vect-R
                     /note="3799 59.3661196848116"
                     /note="color: black; sequence: TTCCTGCCCGACCTTGGT; added: 
                     2020-10-13"
     primer_bind     1222..1248
                     /label=bpk1520-Nme2_INS_R
                     /note="5322 65.1268808195959"
                     /note="color: black; sequence: AAGGTCGGGCAGGAAGAGGGCCTATTT;
                     added: 2020-10-13"
     primer_bind     1222..1248
                     /label=bpk1520-peg-Ins-F
                     /note="3800 65.1268808195959"
                     /note="color: black; sequence: AAGGTCGGGCAGGAAGAGGGCCTATTT;
                     added: 2020-10-13"
     primer_bind     1222..1240
                     /label=pegRNA-IF-Vect-F
                     /note="4351 59.4867747046393"
                     /note="color: black; sequence: AAGGTCGGGCAGGAAGAGG; added: 
                     2020-10-13"
     primer_bind     1234..1254
                     /label=CjeCas9 U6 Scaffold KpnI
                     /note="1821 62.3590543711121"
                     /note="color: black; sequence: 
                     ATATGGTACCGAAGAGGGCCTATTTCCCATG; added: 2020-10-13"
     promoter        1237..1477
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     1237..1260
                     /label=U6_gRNA_PCR_ F
                     /note="255"
                     /note="color: black; sequence: GAGGGCCTATTTCCCATGATTCCT"
     primer_bind     1237..1259
                     /label=PscI_U6promoter-F
                     /note="601 65.202124029999993"
                     /note="color: black; sequence: 
                     TTGCTCACATGTGAGGGCCTATTTCCCATGATTCC; added: 2020-10-13"
     primer_bind     1237..1259
                     /label=MluI_U6promotor-F
                     /note="569 69.113093069364893"
                     /note="color: black; sequence: 
                     GCGCACGCGTGAGGGCCTATTTCCCATGATTCC; added: 2020-10-13"
     primer_bind     1237..1257
                     /label=U6-lentiCRISPR2.0-KpnI-F
                     /note="3812 63.7665731152354"
                     /note="color: black; sequence: 
                     TTGGTTAATTAAGGTACCGAGGGCCTATTTCCCATGATT; added: 2020-10-13"
     primer_bind     1237..1256
                     /label=px601-PURO-U6-F
                     /note="3078 64.0626047482581"
                     /note="color: black; sequence: GACGTGCGAGGGCCTATTTCCCATGAT;
                     added: 2020-10-13"
     primer_bind     1237..1255
                     /label=miniVPR-U6_F
                     /note="3076 62.3142601027153"
                     /note="color: black; sequence: AAAAGGCGAGGGCCTATTTCCCATGA; 
                     added: 2020-10-13"
     primer_bind     1250..1269
                     /label=U6 primer 2
                     /note="140"
                     /note="color: black; sequence: CCATGATTCCTTCATATTTG"
     primer_bind     1366..1388
                     /label=new_U6_FOR
                     /note="4987 53.37999224"
                     /note="color: black; sequence: TTTCTTGGGTAGTTTGCAGTTTT; 
                     added: 2020-10-13"
     primer_bind     1408..1427
                     /label=U6 LKO.1 5' primer
                     /note="122"
                     /note="color: black; sequence: GACTATCATATGCTTACCGT"
     primer_bind     complement(1420..1437)
                     /label=U6_seq1 rv
                     /note="526"
                     /note="color: black; sequence: CTTTCAAGTTACGGTAAG"
     primer_bind     complement(1443..1487)
                     /label=BsaI-to-BbsI-R
                     /note="2479 66.2980766160882"
                     /note="color: black; sequence: 
                     AAGACCCGGTGTTTCGTCCTTTCCACAAGATATATAAAGCCAAGAAATCG; added: 
                     2020-10-13"
     primer_bind     complement(1462..1487)
                     /label=M-NM-sgRNA_BbsI_R
                     /note="300 65.463398936363902"
                     /note="color: black; sequence: 
                     CGAAGACCCGGTGTTTCGTCCTTTCCACAAGAT; added: 2020-10-13"
     primer_bind     complement(1464..1486)
                     /label=xCas9_in55_g1_Q5_Rv
                     /note="color: black; sequence: 
                     gcaagtaacggtgTTTCGTCCTTTCCACAAG"
     primer_bind     complement(1464..1485)
                     /label=m_sa_in54g2_tRNA_rv
                     /note="1094 63.172962567398"
                     /note="color: black; sequence: 
                     CTAGAGCAAGGGTGTTTCGTCCTTTCCACAAG; added: 2020-10-13"
     primer_bind     complement(1467..1487)
                     /label=xcas9_in23_g1_Q5_Rv
                     /note="color: black; sequence: 
                     caggtaagccGGTGTTTCGTCCTTTCCAC"
     primer_bind     complement(1467..1486)
                     /label=xcas9_in22_g1_Q5_Rv
                     /note="color: black; sequence: 
                     tttgaggctcGGTGTTTCGTCCTTTCCAC"
     primer_bind     complement(1467..1486)
                     /label=ABE_in54_g2_Q5_Rv
                     /note="color: black; sequence: 
                     ctctgcaaagcGGTGTTTCGTCCTTTCCAC"
     primer_bind     complement(1467..1486)
                     /label=ABE_in54_g1_Q5_Rv
                     /note="color: black; sequence: 
                     tactctgcaacGGTGTTTCGTCCTTTCCAC"
     misc_RNA        1507..1582
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: RIGHT"
     primer_bind     1507..1526
                     /label=T7_sp_in51g1
                     /note="965 63.9"
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGgacaggacatttaatcaaaggttttagagctaGAAAtagc;
                      added: 2020-10-13"
     primer_bind     1507..1526
                     /label=T7_sp_in54g1
                     /note="964 63.5"
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGcgattgatatagctatgtgagttttagagctaGAAAtagc;
                      added: 2020-10-13"
     primer_bind     1507..1525
                     /label=T7_sp_in47g1
                     /note="963 63.9"
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGagaagttctgctctacgtgagttttagagctaGAAAtag; 
                     added: 2020-10-13"
     primer_bind     1507..1525
                     /label=T7_sp_in54g3
                     /note="966 64.8"
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGcaacagttccgtttaagcaggttttagagctaGAAAtag; 
                     added: 2020-10-13"
     primer_bind     1512..1578
                     /label=peg-Scaffold-T
                     /note="3606 67.9411962359722"
                     /note="color: black; sequence: 
                     AGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCA
                     CCGAGTCG; added: 2020-10-13"
     primer_bind     complement(1516..1582)
                     /label=peg-Scaffold-B
                     /note="3607 68.1172071948587"
                     /note="color: black; sequence: 
                     GCACCGACTCGGTGCCACTTTTTCAAGTTGATAACGGACTAGCCTTATTTTAACTTGCT
                     ATTTCTAG; added: 2020-10-13"
     primer_bind     complement(1559..1582)
                     /label=sp_gRNA_scaff_R
                     /note="967"
                     /note="color: black; sequence: gcaccgactcggtgccactttttc"
     primer_bind     complement(1565..1582)
                     /label=392-libBRCA2-Vect_R
                     /note="5148 61.9744710879193"
                     /note="color: black; sequence: GCACCGACTCGGTGCCAC; added: 
                     2020-10-13"
     primer_bind     complement(1567..1589)
                     /label=gRNA_PCR_R
                     /note="256"
                     /note="color: black; sequence: AAAAAAAGCACCGACTCGGTGCC"
     primer_bind     complement(1567..1586)
                     /label=T7-sgRNA-R
                     /note="2084"
                     /note="color: black; sequence: AAAAGCACCGACTCGGTGCC"
     primer_bind     1568..1582
                     /label=universal_peg_libVect__VR
                     /note="4967 56.9608562447852"
                     /note="color: black; sequence: GCACCGACTCGGTGC; added: 
                     2020-10-13"
     primer_bind     complement(1568..1582)
                     /label=universal_peg_libVect__VR
                     /note="4967 56.9608562447852"
                     /note="color: black; sequence: GCACCGACTCGGTGC; added: 
                     2020-10-13"
     primer_bind     1581..1605
                     /label=CAST SaCas9 insert 1 F Q5
                     /note="3664 69.1764312213406"
                     /note="color: black; sequence: 
                     GCCGCACGCTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1582..1597
                     /label=952_Vect_F
                     /note="5226 58.083996956365"
                     /note="color: black; sequence: 
                     TCGTCTTGCTGTCTTTTTTTAAGCTTGG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 1 fwd
                     /note="3348 61.6522904095318"
                     /note="color: black; sequence: 
                     TCATTCATTTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 2 fwd
                     /note="3350 63.6608282625431"
                     /note="color: black; sequence: 
                     ACCCATCTATTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 3 fwd
                     /note="3352 63.6556551591446"
                     /note="color: black; sequence: 
                     TATGGTGAGTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 4 fwd
                     /note="3354 65.2542884887288"
                     /note="color: black; sequence: 
                     TTCCCTCCTTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 5 fwd
                     /note="3356 63.3502840844038"
                     /note="color: black; sequence: 
                     TGGTTTTGTTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 6 fwd
                     /note="3358 65.3285473144163"
                     /note="color: black; sequence: 
                     AACCTGAGGTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CANX in14 Cas12k 7 fwd
                     /note="3360 62.2665098609781"
                     /note="color: black; sequence: 
                     TTACCCTATTTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CAST SaCas9 insert 2 F Q5
                     /note="3666 63.1111184783889"
                     /note="color: black; sequence: 
                     TGGAAAACATTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CAST SaCas9 insert 3 F Q5
                     /note="3668 63.2559139712726"
                     /note="color: black; sequence: 
                     GCTTCTTCATTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1605
                     /label=CAST SaCas9 insert 4 F Q5
                     /note="3670 64.171277386629"
                     /note="color: black; sequence: 
                     CCCATACGATTTTTTTAAGCTTGGGCCGCTCG; added: 2020-10-13"
     primer_bind     1583..1603
                     /label=bpk1520-Nme2_VECT_F
                     /note="5319 56.0938613311724"
                     /note="color: black; sequence: TTTTTTTAAGCTTGGGCCGCT; 
                     added: 2020-10-13"
     primer_bind     1583..1600
                     /label=Vect_F_566-BRCA-libVect
                     /note="5154 59.122677905929"
                     /note="color: black; sequence: 
                     CACAGAATTCTGTTTTTTTTAAGCTTGGGCC; added: 2020-10-13"
     primer_bind     1583..1598
                     /label=392-libBRCA2-Vect_F
                     /note="5153 56.7785323114552"
                     /note="color: black; sequence: 
                     AATGGTCTCAACTTTTTTTTAAGCTTGGG; added: 2020-10-13"
     primer_bind     1583..1598
                     /label=1061_Vect_F
                     /note="5228 57.6229086554695"
                     /note="color: black; sequence: 
                     CCATGGGCATTAATTTTTTTAAGCTTGGG; added: 2020-10-13"
     primer_bind     1583..1598
                     /label=exon18_peg_libVect__VF
                     /note="4966 57.0216708241011"
                     /note="color: black; sequence: 
                     TGCAGCAGATATTTTTTTTTAAGCTTGGG; added: 2020-10-13"
     primer_bind     1583..1597
                     /label=1186_Vect_F
                     /note="5232 57.9742787294911"
                     /note="color: black; sequence: 
                     GGCTGCCTTTCATTTTTTTTAAGCTTGG; added: 2020-10-13"
     primer_bind     1583..1597
                     /label=Vect_F_10del-BRCA-libVect
                     /note="5156 57.3062191730626"
                     /note="color: black; sequence: 
                     CGATTGGTCAGGTTTTTTTTAAGCTTGG; added: 2020-10-13"
     primer_bind     1590..1605
                     /label=bpk1520-peg-Vect-F
                     /note="3798 59.1765848533689"
                     /note="color: black; sequence: AAGCTTGGGCCGCTCG; added: 
                     2020-10-13"
     primer_bind     complement(1650..1670)
                     /label=BPK2660-U6-R
                     /note="3075 68.8520599859421"
                     /note="color: black; sequence: 
                     AGGCCCTCGCCTTTTTACGGTTCCTGGCC; added: 2020-10-13"
     primer_bind     complement(1671..1685)
                     /label=Fragment.REV (2)
                     /note="2489 72.0018483055393"
                     /note="color: black; sequence: 
                     CGCTACAGGGCGCGTAACGCCAGCAACGCG; added: 2020-10-13"
     primer_bind     complement(1678..1701)
                     /label=ILMNA-LIBVECT-R
                     /note="4475 70.9052260760041"
                     /note="color: black; sequence: 
                     GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGCGGAGCCTATGGAAAAACGCCAG; 
                     added: 2020-10-13"
     primer_bind     complement(1678..1700)
                     /label=sequencing_AAV_F
                     /note="341"
                     /note="color: black; sequence: CGGAGCCTATGGAAAAACGCCAG"
     rep_origin      complement(1686..2274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(1738..1757)
                     /label=ori fw
                     /note="507"
                     /note="color: black; sequence: GTCGGGTTTCGCCACCTCTG"
     CDS             complement(2445..3305)
                     /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: 2445 .. 3236 / #ccffcc
                        2: 3237 .. 3305 / #ccffcc / signal sequence
                     Cleavage site after base 3237"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2564..2585)
                     /label=AmpR_No_BsaI_F
                     /note="1649 64.78606981"
                     /note="color: black; sequence: GCCGCGGTATCATTGCAGCACTGG; 
                     added: 2020-10-13"
     primer_bind     2590..2608
                     /label=AmpR_No_BsaI_R
                     /note="1648 64.27511556"
                     /note="color: black; sequence: TTCCACGCTCACCGGCTCCAG; 
                     added: 2020-10-13"
     promoter        complement(3306..3410)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacgtcgcta gctctagagg gcctgaattc tcgaggcgtt gacattgatt attgactagt
       61 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      121 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      181 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      241 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      301 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      361 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      421 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      481 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      541 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      601 tgggaggtct atataagcag agctctctgg ctaactaccg gtgccaccat ggccatgcct
      661 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
      721 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
      781 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
      841 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
      901 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
      961 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     1021 ccctctggtt atgtgtggga gggctgagaa ttcatcgata ccgtcgagat ctaacttgtt
     1081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1201 ctggaaattc gatccggtac caaggtcggg caggaagagg gcctatttcc catgattcct
     1261 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1321 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1381 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1441 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cgattaatgc
     1501 gtctccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     1561 aaaagtggca ccgagtcggt gcttttttta agcttgggcc gctcgaggta cctctctaca
     1621 tatgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3421 cgaaaagtgc cacct
//
