LOCUS       sequence-215940-        4995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4995)
  TITLE     Ramirez Alvarado plasmids
REFERENCE   2  (bases 1 to 4995)
  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"
COMMENT     Sequence Label: pET12a-HAL
FEATURES             Location/Qualifiers
     source          1..4995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(521..539)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      535..582
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(948..953,954..970,971..976))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1003..1021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1089..1193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1194..1262,1263..2054)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1412..1431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2225..2813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2714..2733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2967..2984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2999..3138
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3064..3083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3224..3246
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3240..3431)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4893..4912
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcgggcga aactcctagg aatagtcctg acaaccccta
      121 tcgcgatcag ctcttttgcg tcgacggaca tccagatgac ccagtctcca tcttccctgt
      181 ctgcatctgt aagagacaga gtcaccatca cttgccgggc aagtcagacc attagcaact
      241 ttttaaattg gtatcagcag aaaccaggta aagcccctaa gctcctggtc tatgctgcaa
      301 ccagtttgca aactggggtc ccctcaggtt tcagtggcaa tggatcgggg acagagttca
      361 ctctcaccat cagcagtctc caacctgagg attttgcaag ttactactgt caacagagtt
      421 acagtacccc gtggacattt ggccaaggga ccaaggtgga aatcaaataa ggatccggct
      481 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      541 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      601 tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag
      661 tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg
      721 tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat
      781 gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat
      841 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
      901 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
      961 gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat
     1021 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1201 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1261 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1321 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1381 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1441 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1501 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1561 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1621 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1681 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     1741 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1801 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1861 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1921 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1981 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2041 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2341 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2581 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2881 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2941 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3001 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3061 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3121 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3181 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3241 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3301 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3361 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3421 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3481 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3541 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     3601 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     3661 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     3721 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     3781 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     3841 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     3901 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     3961 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     4021 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     4081 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     4141 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     4201 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     4261 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     4321 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     4381 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     4441 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     4501 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     4561 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4621 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4681 agggcatcgg tcgatcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     4741 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     4801 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     4861 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     4921 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     4981 tcgatcccgc gaaat
//
