LOCUS       sequence-482913-        4607 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  Template for in vitro transcription of AmNeonGreen mRNA using SP6 
            polymerase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  AUTHORS   n/a
  TITLE     Aiptasia Genetic Tools Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCR182
FEATURES             Location/Qualifiers
     source          1..4607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        879..897
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     879..896
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1686..1705)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1687..1705)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1710..1844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1734..1753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1788..1807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1961..1981)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1961..1979)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2000..2016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2000..2016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2013..2035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2024..2040
                     /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)."
     promoter        complement(join(2048..2054,2055..2072,2073..2078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2093..2114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2231..2248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2402..2990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2482..2501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3161..3952,3953..4021))
                     /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     3784..3803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4022..4126)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4152..4607)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4289..4310)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4501..4520
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gtcgatcgac cgccaattca atatggcgta tatggactca tgccaattca
      121 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
      181 atatggtgga tctggacccc agccaattca atatggctat tggccaggtt caatactatg
      241 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
      301 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
      361 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
      421 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
      481 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
      541 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
      601 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
      661 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
      721 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
      781 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
      841 gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata
      901 caagctactt gttctttttg caggatccca tcgattcgaa ttcgtcgaca tggtgagcaa
      961 gggcgaggag gataacatgg cctctctccc agcgacacat gagttacaca tctttggctc
     1021 catcaacggt gtggactttg acatggtggg tcagggcacc ggcaatccaa atgatggtta
     1081 tgaggagtta aacctaaagt ccaccaaggg tgacctccag ttctccccct ggattctcgt
     1141 ccctcatatc gggtatggct tccatcagta cctaccctac cctgacggga tgagtccttt
     1201 ccaggccgct atggtagatg gctccggata ccaagtccat cgtacaatgc agtttgaaga
     1261 tggtgcctcc cttactgtta actacagata cacctacgag ggaagccaca tcaaaggaga
     1321 ggcccaggtg aaggggactg gtttccctgc tgacggtcct gtgatgacca acagtctcac
     1381 cgctgcggac tggtgcagga gtaagaagac ttaccccaac gacaaaacca tcatcagtac
     1441 ctttaagtgg agttacacca ctggaaatgg caagagatac cgtagcactg cgaggaccac
     1501 ctacaccttt gctaagccaa tggcggctaa ctatctcaag aaccagccta tgtacgtgtt
     1561 ccgtaagacg gaactcaagc actccaagac cgagctcaac ttcaaggagt ggcaaaaggc
     1621 ctttaccgat gtgatgggca tggacgagct gtacaagtag aattcaaggc ctctcgagcc
     1681 tctcgcccta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     1741 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1801 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1861 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggcgcgcc
     1921 gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg
     1981 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2041 ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga
     2101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4141 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4201 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4261 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4321 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4381 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4441 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4501 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4561 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgt
//
