LOCUS       Exported                7581 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  KO of PCC 11901 psbA2 gene/expresses YFP under control of inducible 
            cLac143 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pSW039
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  TITLE     Unprecedented biomass and fatty acid production by the newly 
            discovered cyanobacterium Synechococcus sp. PCC 11901
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7581)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7581
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     110..131
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(198..209)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(222..251)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     protein_bind    300..316
                     /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)."
     protein_bind    401..420
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(1191..1208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1191..1207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1200..1222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1416..1435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1535..1557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1595..1613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1681..1785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1786..2646
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2004..2023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2817..3405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3306..3325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3559..3576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3693..3714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3729..3759
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3767..3783
                     /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)."
     primer_bind     3772..3794
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3791..3807
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3791..3807
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4823..5614
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(5663..6745)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDFSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6707..6726
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      6907..6954
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6950..6968
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(7038..176)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7083..7104)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     7432..7451
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
       61 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
      121 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatcgag
      181 gctgctgcct gcatatgacg accttcgata tggccgctgc tgtgatgatg atgatgatga
      241 tgatgatgat ggcccatgga attaatctcc tacttgactt tatgagttgg tgtgtgaaat
      301 tgttatccgc tcacaattcc acacatttat acaacggtga tgggacttat gtcaattaca
      361 tcttgttaat tttattcctg cttttttgtt aagaattccg aattgtgagc gctcacaatt
      421 cgtttggaga tgtccagagg ggagtcaatt gcatgtacca gtttaaagta ctggggacta
      481 agcaaatcgc tcaatcgatt tattgcttaa tcactttttg attatatagc accgttccta
      541 aaaaagacaa ggcttgcgga aactaaagct gcaaataatc ttttctgtcc gtttcttgag
      601 aggcgatcgc cccattacac taaagtcatt cccactacgc agcaccatgt caaccctgtt
      661 caccagcatc gccaatcgcc tccaaaacgt ttttaaccta gaggttatcg gtgaccaaat
      721 tgcccggggc ctgatcaacc tcgtcgttgc cctcattacc ttcgcagtgt tctactgcct
      781 ttggaagatt gttgatgtgc ccctctatgc atccttcaaa aaatcaaaac tcgataaaac
      841 ctccgccagc ttcttaagga ccctcgttcg ctacgccatt ttgacctttg gcgcgatcca
      901 agccctcaat gcagttggga ttaacatggc cgcaatcgtc accagcctgg gacttgtggg
      961 attaacggtc ggctttgctg cccgggatgc cctttctaac ttgatttctg gactgctcat
     1021 tttttgggat cggccctttg tgattgatga cctcgtggaa attaatggtc tctatggccg
     1081 cgtcgaaaca atcaccctaa ggagcacgcg cattgtgacc gttgatggcc ggatgttagc
     1141 ggtgcctaat tctgtcgtga tcaattcaac ggtgatctcc tacactaatt actggccgtc
     1201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     1261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1321 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     1381 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     1441 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     1501 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     1561 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     1621 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1801 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1861 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1921 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1981 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2041 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2101 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2161 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2221 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2281 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2341 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2401 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2461 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2521 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2581 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2641 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2701 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3481 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3541 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3601 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3661 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     3721 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     3781 caatttcaca caggaaacag ctatgactgg caggcgagac aactgctttc gacattaatg
     3841 cggcggtagt agtggaggcg atcgccttgg atttggctat agctcaggag ttcggggtgt
     3901 tgcgtaaatt tggcgatcgc catggtttcc tggagatttt gcggggcatg gtctggattg
     3961 cgatattttt tagccacctg cttgacttgc caaccattgt cttgactcag ttgcctcgct
     4021 ttcatcccca ctggcttaca aacttcctgc atggtttgcg gcgtgatgtc tgcagtcggg
     4081 tcaatggttt tggccaactc agagcgcatt gggtcaagat cttcgatggc ctgcacggcc
     4141 cgagcaaaat ccgggggatt gggggctgct tgcacaggga caatgggcca aaccaaggtg
     4201 gcgatcgcca aggcactaat gagaatgccg gtcagaaaat gtgaaaactt ttgaccactg
     4261 ctcatttgca cattgaatgg tttgttttta agaataaaac gcaaaatctt cattgtattt
     4321 tttccatttt ttattcaata gtttagtcaa tttctcagga aaaattcgct ttttaggatt
     4381 gataattaaa tcttcttaaa atccaagaaa agcctctact tgaatggcag aggctaagag
     4441 catttagagt atatttttcg ttcaggattc agagcaaacc aagattattc aaagcaatca
     4501 tcatgccact gcaatcccaa attcaaactt ccagaaacgg atgaaggcac gaacccagtg
     4561 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     4621 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     4681 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     4741 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     4801 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     4861 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     4921 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     4981 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     5041 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     5101 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     5161 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     5221 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     5281 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     5341 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     5401 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     5461 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     5521 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     5581 gaaaggcgag atcaccaagg tagtcggcaa ataaaaacca cacgcacaat tcccttaaaa
     5641 agcaacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5701 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     5761 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5821 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5881 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5941 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     6001 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     6061 tttgttgaaa accggacatg gcactaaagt cgccttcccg ttccgctatc ggctgaattt
     6121 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     6181 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     6241 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     6301 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     6361 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     6421 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     6481 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     6541 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     6601 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     6661 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     6721 catcgtataa cgttactggt ttcacattca ccaccatgaa ttgaccctcc tccggctagc
     6781 attgtaccta ggactgagct agctgtaaat gcgccattcg atggtgtccg ggatctcgac
     6841 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6901 tctagacaaa aaacccgtca agacccgttt agaggcccca aggggttatg ctagttattg
     6961 ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatcctg
     7021 ccaaagcacc gccctcgtta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     7081 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     7141 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     7201 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     7261 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     7321 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     7381 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     7441 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     7501 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg caggccgtag
     7561 ccgaaggtgg tcacgagggt g
//
