pWY031 pAM_LplA-FLAG

LOCUS       pWY031 pAM_LplA-FLAG              5268 bp DNA     circular SYN 03-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Sep 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    61..364
                     /label=oriT
                     /label=nonstandard type: ori_tra
                     /note="oriT - streamlined origin of transfer from RP4 for 
                     conjugal transfer. PCR amplified from pRL692 (GenBank: 
                     AF424805)"
                     /note="Babic et al (2008) Research in 
                     Microbiology159:545-549; Koksharova and Wolk (2002) J. 
                     Bacteriol. 184:5524-5528"
                     /note="/id=CVP062"
     misc_feature    414..1305
                     /label=S7942NSIII-LA
                     /label=nonstandard type: recombination
                     /note="Left arm of the homologous recombination site 
                     NS3into the chromosome of Synechococcus elongatus PCC7942 -
                     from pHN1_lacUV5, GenBank: NC_007604, HM026758"
                     /note="Niederholtmeyer et al. Appl. Environ. Microbiol. 
                     76:3462-3466"
                     /note="/id=CVP095"
     primer_bind     1080..1099
                     /label=JC29-NS3-F
     primer_bind     1080..1099
                     /label=NS3-F
     misc_feature    1306..1311
                     /label=NheI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR004"
     misc_feature    1312..1332
                     /label=GC
                     /label=nonstandard type: CV-adaptor
                     /note="Adaptor for the CyanoVECTOR assembly 
                     scheme,flanking: BHR replicon and Neutral sites"
                     /note="/id=CVP005"
     misc_feature    1333..1338
                     /label=NheI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR004-1"
     terminator      complement(1348..1374)
                     /label=trpA T
                     /note="trpA terminator from Lorist6 (GenBank: X98450), 
                     onent shorter than reference"
                     /note="Gibson et al. (1987) Gene 53:283-286"
                     /note="/id=CVP098"
     misc_feature    complement(1382..1915)
                     /label=aacC1-Gm
                     /label=aacC1
                     /label=nonstandard type: ORF_antR
                     /note="Confers gentamycin resistance - gentamicin 
                     acetyltransferase - from pDAH346"
                     /note="Nair et al. (2001) J Bacteriol. 183:1740-1747"
                     /note="/id=CVP016"
     misc_feature    1623
                     /label=snp
     misc_feature    complement(1916..2250)
                     /label=PaacC1
                     /label=nonstandard type: promoter_antR
                     /note="Upstream region of aacC1 ORF incl. promoter and RBS-
                     from pDAH346"
                     /note="Nair et al. (2001) J Bacteriol. 183:1740-1747"
                     /note="/id=CVP078"
     primer_bind     2111..2130
                     /label=JC13-Verify-hly-F
     misc_feature    2251..2256
                     /label=AgeI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR001"
     misc_feature    2257..2277
                     /label=C2G
                     /label=nonstandard type: CV-adaptor
                     /note="Adaptor for the CyanoVECTOR assembly 
                     scheme,flanking: Antibiotic resistance markers; 
                     Expression/reporter cassette, promoter-reporter module, 
                     custom sequence"
                     /note="/id=CVP006"
     misc_feature    2278..2283
                     /label=AgeI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR001-1"
     terminator      2303..2331
                     /label=lpp T
                     /note="lpp T - lpp terminator"
                     /note="Cao and Sarkar (1992) PNAS 89:7546-7550"
                     /note="/id=CVP099"
     primer_bind     2347..2392
                     /label=pPsbA1-3F
                     /note="NS3 Vtctor 26bp (including SwaI 6bp)+ pPsbA1 20bp"
     misc_feature    2365..2368
                     /label=SwaI
                     /label=nonstandard type: restrict_site
                     /note="NIL"
                     /note="/id=CVR006"
     promoter        2373..2530
                     /label=pPsbA1
     primer_bind     complement(2505..2532)
                     /label=NS3-BB-R1
     primer_bind     complement(2511..2571)
                     /label=JC17-pPsbA1-1R
                     /note="secrection signal 20bp + EcoRI 6bp + pPsbA1 20bp"
     primer_bind     2511..2557
                     /label=lplA-ns3-F1
     primer_bind     2511..2557
                     /label=ns3-lplA-F
     primer_bind     complement(2511..2539)
                     /label=pPsbA1-3R
                     /note="hly-flag 26bp (including EcoRI 6bp) + pPsbA1 20bp"
     primer_bind     2531..2557
                     /label=LplA-6803-F1
     misc_feature    2531..2532
                     /label=EcoRI
     primer_bind     2535..2557
                     /label=LplA-F2
     primer_bind     2536..2566
                     /label=NcoI-F
     CDS             2537..3553
                     /codon_start=1
                     /label=LPLA
                     /translation="MGSTLRLLISDSYDPWFNLAVEECIFRQMPATQRVLFLWRNADTV
                     VIGRAQNPWKECNTRRMEEDNVRLARRSSGGGAVFHDLGNTCFTFMAGKPEYDKTISTS
                     IVLNALNALGVSAEASGRNDLVVKTVEGDRKVSGSAYRETKDRGFHHGTLLLNADLSRL
                     ANYLNPDKKKLAAKGITSVRSRVTNLTELLPGITHEQVCEAITEAFFAHYGERVEAEII
                     SPNKTPDLPNFAETFARQSSWEWNFGQAPAFSHLLDERFTWGGVELHFDVEKGHITRAQ
                     VFTDSLNPAPLEALAGRLQGCLYRADMLQQECEALLVDFPEQEKELRELSAWMAGAVR"
     primer_bind     2537..2557
                     /label=LplA-35s-F
     primer_bind     2540..2560
                     /label=LplA-F1
     primer_bind     complement(2924..2943)
                     /label=LplA-seq-R1
     primer_bind     complement(3254..3280)
                     /label=LplA-N-R
     misc_feature    3281..3553
                     /label=C domain
     primer_bind     3281..3304
                     /label=lplA-C-F
     primer_bind     complement(3534..3553)
                     /label=LplA-R1
     primer_bind     complement(3535..3553)
                     /label=LplA-his R
     primer_bind     complement(3536..3553)
                     /label=XhoI-R
     primer_bind     complement(3540..3583)
                     /label=NS3-LPLA-R2
     primer_bind     3555..3581
                     /label=NS3-BB-F2
     misc_feature    3560..3625
                     /label=3×Flag
     primer_bind     3560..3581
                     /label=NS3-BB-F2-
     primer_bind     complement(3601..3635)
                     /label=Lpla-6803-R2
     primer_bind     complement(3602..3635)
                     /label=LplA-6803-R1
     primer_bind     complement(3609..3706)
                     /label=hly-flag-R
                     /note="NS3 Vector 26bp(including HindIII) + rrnB T1 46bp + 
                     EcoR32I 6bp + hly-Flag 20bp"
     primer_bind     3629..3658
                     /label=NS3-BB-F1
     misc_feature    3635..3680
                     /label=rrnB T1
     misc_feature    3687..3692
                     /label=XbaI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR007-1"
     misc_feature    3693..3713
                     /label=G5C5
                     /label=nonstandard type: CV-adaptor
                     /note="Adaptor for the CyanoVECTOR assembly 
                     scheme,flanking: Expression/reporter cassette, 
                     promoter-reporter module, custom sequence ; Cyanobacterial 
                     replicons"
                     /note="/id=CVP002"
     misc_feature    3714..3719
                     /label=XbaI
                     /label=nonstandard type: restrict_site
                     /note="/id=CVR007"
     misc_feature    3720..4521
                     /label=S7942NSIII-RA
                     /label=nonstandard type: recombination
                     /note="Right arm of the homologous recombination site 
                     NS3into the chromosome of Synechococcus elongatus PCC7942 -
                     from pHN1_lacUV5, GenBank: NC_007604, HM026758"
                     /note="Niederholtmeyer et al. Appl. Environ. Microbiol. 
                     76:3462-3466"
                     /note="/id=CVP093"
     primer_bind     complement(3796..3815)
                     /label=JC30-NS3-R
     primer_bind     complement(3796..3815)
                     /label=NS3-R
     misc_feature    4532..5205
                     /label=ori-pBR322
                     /label=nonstandard type: ori_rep
                     /note="Origin of replication of pBR322"
                     /note="/id=CVP059"
ORIGIN
        1 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
       61 ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc
      121 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
      181 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
      241 tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa
      301 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
      361 gatccctgac gggcttgtct gctcccgcca tccgcttaca gacaagctcc gcgcgtcgtg
      421 atgtcgaaac ccaagccacc ctcagaggtg aaggccgctt cgagcacatc gggaagcgtg
      481 tcgacgacgg tgctcggtgc ctcgggtaag agccagatag atgcggtgac gttcaccgtc
      541 gcgatcgtgg cgggaaccac ggtcactgcg tccgttacaa ctcgcacgtc gtcggccagc
      601 acgactgact cgacggcttc gattagcccg ggagaggcaa ggccatcagg ctcggtagac
      661 agaatgctga ttaacacctc gccgggagca gggctgctca cagccgcgtc cttcacccgt
      721 gggtcagccg tcagcgcttg atagcggtac caggccgcac cgcctgcggt cgagctgccc
      781 ttgatccgct cgatggtgcg atcgcgaagc tcgccatccg gctcattcgg ctgccgcacc
      841 acgccgtaga acgcggcaag gttgtcgagg tcggggccgt ttgagtagcg aagcagtgtc
      901 gcaagaagtg cgtcgttgat ccgctgacgc aggatcagct cgcgggctgc gcagacctcc
      961 agcagcttga tgaccgggtc ggattcgagg atggcggtgt agctggcgtc acgcgatcgc
     1021 aggtcgtcga tcaagtcctg caggatcagt tcaaagtcca gcgcttcgat gatggtgggc
     1081 gcgggaatcg tagcaaagtc aagaacggtc atgagacgac taagccctcc agcgtgatac
     1141 gcctgccctc ggggatgtag tagccgatca ggttcagctc gacttgacca gctgcgctgg
     1201 ctgagacgat gcgaaccttc tccagcttca gccgtggctc ccagcgatcg agcgcttcag
     1261 ctgtggccgc caccaggtca acgatgaggg actggttgat cggtcgctag catcgcgcgc
     1321 gcgcgcgcgg atgctagcgg cgcgccaaaa aaaagcccgc tcattaggcg ggctggccgg
     1381 cttaggtggc ggtacttggg tcaatatcaa agtgcatcac ttcttcccgt atgcccaact
     1441 ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag
     1501 caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc
     1561 tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa
     1621 acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa
     1681 gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt
     1741 gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg
     1801 atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta
     1861 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg
     1921 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
     1981 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     2041 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     2101 tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac
     2161 ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg
     2221 gcgaacgagc gcaaggtttc ggtctccacg accggtatcc cgccgccgcc gccggatacc
     2281 ggttaatagt acctgtgaag tgaaaaatgg cgcacattgt gcgacatttt tttctcgagc
     2341 aggtagtcgg ccaataaccc agggatttaa atgatcgctc taaacattac ataaattcac
     2401 aaagttttcg ttacataaaa atagtgtcta cttagctaaa aattaagggt tttttacacc
     2461 tttttgacag ttaatctcct agcctaaaaa gcaagagttt ttaactaaga ctcttgccct
     2521 ttacaacctc gaattcatgg gctccacatt acgcctgctc atctctgact cttacgaccc
     2581 gtggtttaac ctggcggtgg aagagtgtat ttttcgccaa atgcccgcca cgcagcgcgt
     2641 tctgtttctc tggcgcaatg ccgacacggt agtaattggt cgcgcgcaga acccgtggaa
     2701 agagtgtaat acccggcgga tggaagaaga taacgtccgc ctggcgcggc gcagtagcgg
     2761 tggcggcgcg gtgttccacg atctcggcaa tacctgcttt acctttatgg ctggcaagcc
     2821 ggagtacgat aaaactatct ccacgtcgat tgtgctcaat gcgctgaacg cgctcggcgt
     2881 cagcgccgaa gcgtccggac gtaacgatct ggtggtgaaa accgtcgaag gcgaccgcaa
     2941 agtctcaggc tcggcctatc gcgaaaccaa agatcgcggc ttccaccacg gcaccttgct
     3001 actcaatgcc gacctcagcc gcctggcaaa ctatctcaat ccggataaaa agaaactggc
     3061 ggcgaaaggc attacgtcgg tacgttcccg cgtgaccaac ctcaccgagc tgttgccggg
     3121 gatcacccat gagcaggttt gcgaggccat aaccgaggcc tttttcgccc attatggcga
     3181 gcgcgtggaa gcggaaatca tctccccgaa caaaacgcca gacttgccaa acttcgccga
     3241 aacctttgcc cgccagagta gctgggaatg gaacttcggt caggctccgg cattctcgca
     3301 tctgctggat gaacgcttta cctggggcgg cgtggaactg catttcgacg ttgaaaaagg
     3361 ccatatcacc cgcgcccagg tgtttaccga cagcctcaac cccgcgccgc tggaagccct
     3421 cgccggacga ctgcaaggct gcctgtaccg cgcagatatg ctgcaacagg agtgcgaagc
     3481 gctgttggtt gacttcccgg aacaggaaaa agagctacgg gagttatcgg catggatggc
     3541 gggggctgta aggctcgagg attacaagga tcatgatggt gattataagg atcacgacat
     3601 cgactataaa gacgatgatg acaaataaga tatccaaata aaacgaaagg ctcagtcgaa
     3661 agactgggcc tttcgtttta aagctttcta gaatcggggg cccccggggg gattctagac
     3721 gtgagccgta gtcccgtcgc cagacgcggg tgcccacggg cgtcgtcagg atgtccgtaa
     3781 ttgactgccg gaggtggtca atgcccttca gctccttgcc actgtcacgg ctcatgcctc
     3841 gggtcattag tcgcccgctc cggtatcttc actggcttcg atgattgccg ccccgcagct
     3901 gcagaggtca ccgatccgag cagtcggcct ctggttggta aagaccgtgc gactgccggt
     3961 gatgatcgtg ttcaagccat gcagggggca ggcgtgaagg tcatcctttc gggccacggg
     4021 gcggctgttg acgaaggtgt cgtcgctccc ggtgatgatg atcccgccgt gatcggtcac
     4081 gtcgtttagt cgagcgatgc ctggcgtcgt agtcacgggt ttaggtcaat acgacttgcg
     4141 gtcactgtaa cgttgccctc ggcggtcacg ttaacgtcgc cttgggcttc gacttgcgcc
     4201 tcctgcacaa ggatcacaat ccgtccttgg gctgcggtga ggtcgatctt gtactcatgc
     4261 gcttcgcggt cgtactggat gattgagtca tcctcgaact gcgtcttttg gatcgtttct
     4321 ttgtcctcga tctgggggta gtcagtcgag aacgcgccgg gcatcgcgaa gccctgactg
     4381 atctcgccgg agggggccat cacgacgacg gcctcaccga cctcgggcgc ccaccagaac
     4441 cgatccttgc ccgctcgctg cgtgagccac ggaatccagt cagtgaggag cagcgcctcg
     4501 ccgctctcct cgtcttcctc gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4561 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4621 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4681 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4741 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4801 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4861 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4921 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4981 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5041 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5101 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5161 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5221 atcccctgat tctgtggata accgtattac cgcctttgag tgagctga
//