LOCUS       sequence-383139-        6269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAD322-MycaTnsD, arapBAD promoter expressing cyanobacterial 
            tRNA-Leu gene targeting TnsD protein of McCAST (Tn7575)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6269)
  AUTHORS   Hsieh SC, Peters JE
  TITLE     Discovery and characterization of novel type I-D CRISPR-guided 
            transposons identified among diverse Tn7-like elements in 
            cyanobacteria.
  JOURNAL   Nucleic Acids Res. 2023 Jan 25;51(2):765-782. doi: 
            10.1093/nar/gkac1216.
  PUBMED    36537206
REFERENCE   2  (bases 1 to 6269)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2023-01-25- 25"; volume: "51"; issue: 
            "2"; pages: "765-782"
COMMENT     Sequence Label: pOPO646
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(425..1303)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1330..1614
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1537..1556
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3044..3061)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3214..3300
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3392..3419
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3438..3529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3530..3598,3599..4390)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3748..3767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4432..4887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4519..4538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4729..4750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4998..5586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5487..5506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5740..5757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5772..5912
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5838..5857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5998..6020
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6014..6205)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
       61 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
      121 ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc
      181 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
      241 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
      301 gtgataaact accgcattaa agctagctta tcgatgcata atgtgcctgt caaatggacg
      361 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
      421 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
      481 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
      541 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
      601 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
      661 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
      721 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
      781 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
      841 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
      901 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
      961 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     1021 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     1081 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     1141 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     1201 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     1261 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     1321 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
     1381 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
     1441 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
     1501 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
     1561 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
     1621 ttttttgggc tagcaggagg aattcaccat gacttcaccc attgctgaac attgggatct
     1681 catatatcct gctagtccac ctcgcagtcg tctttacagt ttgcaaccga taggagtcgg
     1741 aactccttgg gttgagagct tgactggata catggctcgg ctagcagaag cacactgcgt
     1801 ctcgcctgga gatttaattg ccaatgaact taaatctttg gctagcaaat ctcacggaaa
     1861 aagctactta catcagacaa gtagctctac agaaagtcta aatagcaccg gacgtatggc
     1921 attggaattg gttcaagcac tggagtatca aaccctacgt ctcgatctca gatatctaac
     1981 cctcttgtcg tgggctgaca ttttgccgcc gaaaggttta ttacgacgta tccgtgcttg
     2041 gtgtcctagt tgctacaatg agtggcaaca caatcagaaa cctatctatg agccactcct
     2101 atgggcacta gaaacggtct caatctgctt aaaacatcgg cagcaactgc gctcatgttg
     2161 cccaaaatgc caacgccaac tacccgcatt ggcgtggcat tctaaacctg gatactgttc
     2221 tagatgctac caatggttag gtacaactct tactccggaa gtagctacct taacaacaaa
     2281 tcccgacgaa ctaaaatggc aaagatgggt tttagagaat gtagggaaat tactcgaaac
     2341 tgctcctagc ttattaaaag cacctactag ggagcgaatt tctcgttcgc tctctgtttg
     2401 tatctcgcgc accgtcgaag ggaatattgc tgagttcgct cgtcaattag gaacacacaa
     2461 aagtgcagta tggcagtgga agtcgggcaa agccttgcct caattgtcaa tgcttctcaa
     2521 aatctgtcaa attttagaga tagacctact aaatttcttg tttaatccag aatctttaac
     2581 taacacacgc gattcgttag ccattttggt aaagcgagac ttcccttctc aaaaacctaa
     2641 acaacagctt gatactgatc aagtacaaga attcttgcaa tctgccctta tagaagaacc
     2701 tcctccaact atgaaggctg ttgcccaaac gttgggacac aattcccgat cgctccgacg
     2761 ccgatttcca gaactgtgtt ctgctatctc ggctcggtac ttaaattacc gagatcaggt
     2821 tcgtgctgca aaagttcaag agtcttgcca agaggtacgt cagattgctc tcaaacttta
     2881 tggtgaaggt attgatccta cacgcagcta catatccagg tacctaagca agccagcata
     2941 ttttcgtgaa cctgctgttg tcgccgtact agagtcaact cgacaagaac taggacttaa
     3001 aaattcttaa tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     3061 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3121 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3181 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3241 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3301 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     3361 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     3421 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3481 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3541 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3601 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3661 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3721 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     3781 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3841 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3901 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3961 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4021 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4081 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4141 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4201 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4261 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4321 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4381 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     4441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4621 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4741 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4801 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4861 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     4921 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5101 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5161 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5221 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5281 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5341 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5401 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5461 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5521 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5581 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5641 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5701 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5761 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5821 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     5881 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     5941 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6001 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     6061 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     6121 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     6181 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     6241 gggtaatgat accgatgaaa cgagagagg
//
